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Comm. Appl. Biol.

Sci, Ghent University, 76/2, 2011 i

VOL 76/2 1-99 (2011)

COMMUNICATIONS IN AGRICULTURAL AND APPLIED


BIOLOGICAL SCIENCES

Formerly known as

MEDEDELINGEN FACULTEIT LANDBOUWKUNDIGE EN


TOEGEPASTE BIOLOGISCHE WETENSCHAPPEN

Publishers
Prof. Guy Smagghe
Prof. Pascal Boeckx
Prof. Peter Bossier
Prof. Walter Steurbaut
Prof. Els JM van Damme
Prof. Niko Verhoest

Editorial adress
Coupure links 653
9000 Gent (Belgium)

Website
http://www.plantpower.eu
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 ii
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 iii

The results published in this book of abstracts are under the full
responsibility of the authors. The organizing committee cannot be held
responsible for any errors in this publication and potential consequences
thereof.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 iv
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 v

PROCEEDINGS

1st international PlantPower Symposium

February 10th, 2011


Ghent, Belgium

FACULTY OF BIOSCIENCE ENGINEERING


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 vi

MEMBERS OF THE ORGANIZING COMMITTEE

Jan Arends, Ghent University


Christine Graveel, Ghent University (Secretary)
Arnd Kuhn, Forschungszentrum Jülich
David Strik, Wageningen University
Bert Hamelers, Wageningen University
Kirsten Steinbusch, Wageningen University (IT)
Tim Lacoere, Ghent University (IT & Graphics)
Nico Boon, Ghent University
Willy Verstraete, Ghent University

MEMBERS OF THE FP7 PLANTPOWER PROJECT GROUP

Scientific partners

Business partners
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 vii

THIS SYMPOSIUM RECEIVED SUPPORT FROM THE EUROPEAN


COMMUNITY’S SEVENTH FRAMEWORK PROGRAMME FP7/2007-2013
UNDER GRANT AGREEMENT NO. 226532.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 viii
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 ix

PREFACE
The fact that micro-organisms can exchange electrons with solid materials
and thus can generate electrical currents which can be used for various
purposes has generated a hype of interest in the last decade. Even more
striking is the fact that growing plants can transfer their metabolites to
micro-organisms and thus, be it indirectly, also produce measurable
electrical currents. This phenomenon constitutes the discussion platform
of this symposium.

The concept of creating an intensive interphase between a higher plant


and its surrounding microbiota is not new. Techniques to empower plants
with rhizospheres or mycorrhiza which are beneficially interacting with the
plant have been the subject of lots of fundamental and applied studies.
Yet, active management of the root microbiome by means of explicit
electrical systems is very novel. Clearly, it is too early to relate the
current explorations to applications in the nearby future. Nevertheless, It
must be clear that the research not only should deal with blue sky
questions about the microbial ecology as such, but should indeed aim to
translate the findings as much as possible to become step stones for
potential practical implementations.

The overall picture is very powerful: the plant as a sustainable capturing


device of light, and the energy transferred via the microbes to generate
’an added value’. We need clever thinking and a dose of serendipity to
make this work properly. Moreover, although the potentials are enormous,
we should not be afraid to be honest about the actual state of the art and
rate of progress.
Overall, we have good confidence that this symposium will be a
challenging ground for direct and lateral constructive interactions.

W. Verstraete
LabMET, Ghent University
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 x

THE PLANTPOWER PROJECT


BERT HAMELERS
Department of Environmental Technology
Wageningen University
Wageningen, the Netherlands

The PlantPower concept is based on the cooperation of plants and micro-


organisms to produce in-situ electricity. Plants take up carbon dioxide and
water and capture light energy. This energy is stored in the chemical bonds
of sugars produced, using carbon dioxide and water. Part of this chemically
stored energy is transferred to the roots of the plants. This energy present
in the root zone can then be captured by the so-called electro-chemical
active bacteria. These organisms are capable to oxidize the organic matter
present in the root zone and transfer the energy rich electrons to an
electrode. The energy carried by the electrons can be used as electrical
energy, after which the electrons react at another electrode with oxygen
to form water.

Figure 1. Schematic of the Plant Power concept.


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xi

The primary advantage of the PlantPower concept is that renewable, clean


electricity can be produced while the facility can be well integrated in the
landscape. The aim of the EU project is to enhance the productivity of the
Plant Power concept such that it becomes competitive with other bio-
energy systems. To achieve this all elements (plant, roots, root microbial
community and fuel cell technology) forming the concept need to be
improved in an integrated fashion. To achieve this goal research groups and
companies with complementary backgrounds have joined forces in this
project to bring this concept forward. Also a spin-off company Plant-e has
early realised to bring forward this technology more commercially.

10
3.2 W/m2 theoretical maximum
Electricity generation

1
0.3 W/m2 biomass electricity
(W/m )
2

0.1

0.01

0.001
2006 2007 2008 2009 2010 2011 2012
Year of experiment

Figure 2. Progress in Power Density of Plant-MFCs. (Data adapted from Strik et al.,
Trends in Biotechnology 29 (1) 2011 & unpublished results PlantPower consortium)

The efforts from all participants have led to a steady progress in the power
density. This progress is depicted in the figure 2. We see a steadily
increasing power density resulting from plant selection and fuel cell
design. The performance of the Plant Power concept can thus reach
already levels comparable to currently employed bio-energy systems.
Considering that we are using only small part of the energy available in the
root zone, we expect further improvements to be realisable up to 3.2
W/m2.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xii

PLANTPOWER – AN "ADVENTURE" PROJECT IN THE


ENERGY THEME
The FET experience

CARLOS SARAIVA MARTINS


European Commission

The challenge for policy-makers in the field of research, technology and


innovation, particularly in the Energy context, is the timely identification
of new directions that have a high potential for significant breakthrough
and may become tomorrow’s energy robust technologies. It is thus
essential to encourage a shift from incremental progress towards more
radical changes.
Under the Energy Theme, beside the classical activities, one is also
addressing in an open and flexible way Future and Emerging Technologies -
FET. The idea is to support research aiming at identifying or further
exploring new scientific and technological opportunities in a given field
and/or in their combination with other relevant areas and disciplines
through specific support for spontaneous research proposals.

Dealing with emerging technologies in an open and flexible way is a FP7


requirement and these calls have proven to be a success. Launching
completely bottom-up calls, based on novel approaches within the overall
goals of the Energy Theme ensured a genuine chance for “emerging needs”
to be funded. There is a strategic need to stimulate a creative spirit in
European research, to provide rewards for “high risk / high impact”
science, to vigorously promote multidisciplinarity on an international
collaborative basis in the European environment.

The publication of such calls minimised the risk of mismatch between


setting priorities and the most meaningful developments in research. If
priorities become too narrowly focussed in terms of detailed scientific
topics, opportunities are missed. The FET calls constituted the natural
obvious answer and PlantPower is a good example.

The research supported has an orientation towards long term innovation


but it is clearly “purpose driven” and not “blue-sky”. The project
objectives are tangible, highly ambitious and challenging. This means
either reaching a clearly defined scientific goal and/or creating a new
basic technology, which in either case has the potential to open up new
fields of enquire and lies well beyond the international state of the art.
So far we have no real success stories from PlantPower (or any other
project). When developing a new technology one often encounters gaps in
understanding that require going back to science to develop new
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xiii

knowledge. A lot of these FET projects will not be able to deliver what
they have promised – that's why due to their high risk character a GO/NO
GO decision at mid-term is implemented. Living plants and bacteria
generating electricity is that possible? Here we have the biggest risk of the
project - is that the concept is not possible at all. But it is worth trying!

From the strategy (and also budget point of view), a balance is needed
between the 2020 time horizon – especially large-scale demonstrations
that can trigger deployment – and the longer term – 2050 and the need for
technology breakthroughs. The feedback loop between science and
technology is a critical part of how progress is made. The more active the
feedback loop, the higher the likelihood of rapid success. This key element
of innovation is at the core of FET projects like PlantPower.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xiv

CONTENT

PREFACE ....................................................................................................... IX

THE PLANTPOWER PROJECT ................................................................................ X

PLANTPOWER: AN “ADVENTURE” PROJECT IN THE ENERGY THEME ............................... XII

PROGRAM .....................................................................................................XVI

SESSION A: PLANTS & RHIZODEPOSITION .................................................................. 1

Keynote: Prof. Dr. Gőnter Neumann ........................................................................ 2

Exploration of key rhizosphere parameters in plant-MFCs ............................................... 7

Non-invasive quantification of root growth via NMR-imaging ........................................... 11

Quantification of exudation of the plant-MFC ............................................................ 15

Nutrient supply influences root exudation and development ........................................... 19

SESSION B: MICROBIAL ASPECTS OF THE RHIZOSPHERE & ANODE .................................. 23

Keynote: Prof. Dr. Michael Friedrich ....................................................................... 24

Microbial Resource Management (MRM): theory and practical tools to exploit


bacterial capabilities ......................................................................................... 27

Comparison of bacterial rhizosphere communities from plant-MFC’s with different


current production by 454 amplicon sequencing ......................................................... 31

Methanotrophic microbiome ................................................................................. 33

SESSION C: MFC TECHNOLOGY ............................................................................ 37

Keynote: Prof Dr. John Greenman .......................................................................... 38

Cathodes for sediment MFCs ................................................................................. 43

Anode materials for sediment MFCs ........................................................................ 47

Autotrophic nitrous oxide removal in bio-electrochemical systems ................................... 51

Year round performance of the flat-plate plant-MFC .................................................... 55


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xv

Electrochemical dechlorination of chlorinated compounds in a Microbial Electrolysis


Cell (MEC) ...................................................................................................... 59

Bacterial mutualism in the mosses roots applicable in Bryophyta-MFC................................ 63

Modified electrodes for more powerful and sustainable plant-MFCs ................................... 67

Coulombic efficiency in a PMFC, effect of exudates, pH and oxygen.................................. 71

Polymer derived carbons and their application in PMFC’s ............................................... 75

Radial oxygen loss decreases available substrate for electrochemically active


bacteria ......................................................................................................... 79

SESSION D: IMPLEMENTATION & IMPACT ................................................................ 83

Keynote: Prof. Dr. Louise Vet ............................................................................... 84

Harnessing solar energy in Bio-PhotoVoltaic (BPV) systems ............................................. 89

Modeling the plant-MFC ...................................................................................... 93

Energetic performance of microbial solar cells ........................................................... 97


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xvi

PROGRAM
st
1 International PlantPower Symposium

Thursday February 10th, 2011


Registration and welcome coffee
(Coupure links 653 – Blok E (ground floor), B-9000 Gent)
08h30-09h00

Opening
(Coupure links 653 – Blok E, room 1.012 B-9000 Gent)
09h00-09h130
Prof. Dr. Willy Verstraete
(Ghent University, Belgium)
Dr. Bert Hamelers
(Wageningen University, The Netherlands)
Dr. Carlos Saraiva-Martins
(European Commission)

Morning session :

Keynote A: Plants & Rhizodeposition


Prof. Dr. Gőnter Neumann
“Rhizodeposition – An Overview”
(Hohenheim University, Germany)
09h30-10h00

Keynote B: Microbial aspects of the Rhizosphere & Anode


Prof. Dr. Michael Friedrich
“The microbial ecology of Electricigenic microorganisms in plant-rhizosphere
based microbial fuel cells”
(University of Bremen)
10h00-10h30

Coffee Break
10h30-10h50
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xvii

Session A & B:
Short oral introduction of Posters
10h50-12h00

Contributed posters A:
Blossfeld et al. (Forschungszentrum Jőlich)
“Exploration of plant functioning and vigour in Plant-MFC”
Blossfeld et al. (Forschungszentrum Jőlich)
“Non-invasive quantification of root growth via NRM-imaging”
Kuijken et al. (Wageningen University)
“Quantification of exudation of the Plant-MFC”
Le Marié et al. (Forschungszentrum Jőlich)
“Influence of the root environment on the root physiology”

Contributed posters B:
Marzorati et al. (Ghent University)
“Microbial resource management (MRM): theory and practical tools to exploit
bacterial capabilities”
Rothballer et al. (HelmholtzZentrum Mőnchen)
“Comparison of bacterial rhizosphere communities from Plant-MFC’s with
different current production by 454-sequencing”

Lunch & Poster Session A & B


(Coupure links 653 – Blok E (ground floor), B-9000 Gent)
12h00-13h30

Afternoon session:

Keynote C: Microbial Fuel Cell Technology


Prof. Dr. John Greenman
“Overview of microbial fuel cells for practical power production”
(Bristol Robotics lab)
13h30-14h00

Keynote D:Implementation and Impact


Prof. Dr. Louise Vet
“Learning from nature: need, challenge and implementation of eco-technology”
(Netherlands institute of Ecology)
14h00-14h30

Coffee Break
14h30-14h50
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 xviii

Session C & D:
Short oral introduction of Posters
14h50-16h00

Contributed posters C:
Desloover et al. (Ghent University)
“Autotrophic nitrous oxide removal in bio-electrochemical systems”
Helder et al. (Wageningen University)
“Year round performance of the flat-plate Plant-MFC”
Hennebel et al. (Ghent University)
“Electrochemical dechlorination of chlorinated compounds in a microbial
electrolysis cell (MEC)”
Hubenova et al. (University of Plovdiv)
“Bacterial mutualism in the mosses roots applicable in Bryophyta-Microbial fuel
cell”
Picot et al. (University of Rennes)
“Modified electrodes for more and sustainable plant-MFC”
Steinbusch et al. (Wageningen University)
“Coulombic efficiency in a PMFC, effect of exudates, pH and oxygen”
Tennisson et al. (MAST Carbon)
“Polymer derived carbons and their application in PMFC’s”
Timmers et al. (Wageningen University)
“Radial oxygen loss decreases available substrate for electrochemically active
bacteria”

Contributed posters D:
Bombelli et al. (University of Cambridge)
“Harnessing solar energy in bio-photovoltaic (BPV) systems”
Strik et al. (Wageningen University)
“Energetic performance of microbial solar cells”

Poster Session C & D


(Coupure links 653 – Blok E (ground floor), B-9000 Gent)
16h00-17h00

Poster Award & Closing Remarks


17h00

Reception
17h30

Traditional Belgium Dinner


(Restaurant ‘t Klokhuys, Corduwanierstraat 65, B-9000 Gent)
19h00
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 1

A:
PLANTS & RHIZODEPOSTION
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 2

KEYNOTE BY PD DR. GÜNTER NEUMANN


Research interests: Plant-nutritional functions of rhizosphere processes,
physiology of root exudation, root exudates and plant-microbial
interactions.

The focus of rhizosphere research in the rhizosphere working group at the


Institute of Crop Science in Hohenheim comprises turnover of
rhizodeposits, nutrients and pollutants in the rooting zone, mechanisms of
root exudation, and perspectives for rhizosphere management. Within the
university, links in teaching and research exist with the departments of soil
science, soil microbiology, crop production, plant breeding and
phytopathology. Special emphasis is placed on international cooperation
networks, reflected e.g. by participation and hosting of four EC-funded
projects on rhizosphere research during the last decade, and by PhD
students and scientists from more than 10 countries as non-permanent or
permanent members of the department.

CV

Studied plant physiology at the “Institute of Botany “, University of


Tübingen (working groupo Prof. D, A, Hager) and at the “Institute of Gene-
Biological Research”, Berlin (working group Prof. Dr.A. Brennicke).
PhD 1992 on Flavonoid biosynthesis in Oenothera

Post Doc at the “Institute of Plant Nutrition”, University of Hohenheim


(working group of Dr. H. Marschner).
Since 2002 Senior Scientist at the “Institute of Plant Nutrition”, Hohenheim
University, Germany - Rhizosphere and Fertilization Working group

2008 Habilitation in Plant Nutrition, Hohenheim University, Germany

Since 2010 Group leader “Rhizosphere working group” at the “Institute of


Crop Science”, “Chair for Nutritional Crop Physiology”, Hohenheim
University, Germany
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 3

RHIZODEPOSITION – AN OVERVIEW

GÜNTER NEUMANN

Institute of Crop Science (340h)


Chair of Nutritional Crop Physiology,
Universität Hohenheim, 70593 Stuttgart, Germany
Corresponding author E-mail: guenter.neumann@uni-hohenheim.de

In higher plants, a substantial proportion (20–60%) of carbon fixed during


photosynthesis can be translocated below ground. Depending on root activity, 15–
60% of this carbon fraction is used for root respiration and is finally released as
CO2. Of the assimilates translocated below ground, up to 70% in perennials and up
to 40% in annual plants enter the soil as organic rhizodeposition, corresponding to
800–4,500 kg carbon ha–1 year–1. This is associated with a concomitant input of
nitrogen ranging between 15 and 60 kg ha–1 year–1 (Fig. 1). However, available
quantitative data on rhizodeposits are highly variable, and strongly influenced by
the methodological approaches, sampling conditions, plant species and genotypes.

Rhizodeposition comprises lysates of sloughed-off cells and tissues resulting from


root turnover (up to 50% of carbon translocated below ground) and root exudates
released from intact root cells. Root exudates can be further subdivided into (1)
diffusates – organic compounds continuously lost from plant roots by diffusion, (2)
root excretions released as metabolic waste products, or (3) secretions with special
functions in nutrient mobilisation, detoxification, plant-microbial signalling and
defense reactions.

Root exudates may comprise 5–10% of the net fixed carbon in soil-grown plants.
The controlled liberation of so-called root border cells, released as living root cells
from the root, also contributes to rhizodeposition to some extent. However, the
related carbon input has been calculated to account only for 1–2% compared with
the carbon fraction released as root exudates (Fig. 1).

The outstanding importance of the rhizosphere for cycling of carbon and nutrients
in soils is illustrated by the fact that organic rhizodeposition, which can account for
30–40% of the total soil organic matter input, is released into the rhizosphere soil,
which comprises only 2–3% of the total soil volume.

Composition, quantity and rhizosphere concentrations of rhizodeposits are highly


variable in space and time. Quality, quantity, solubility and stability of released
compounds in the rhizosphere are strongly influenced by plant-related and
environmental factors: The plant nutritional status, photosynthetic activity, root
morphology, expression of retrieval mechanisms for organic compounds, plant-
developmental stage and genotypic differences are the most important plant
factors with impact on rhizodeposition.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 4

Figure 1. Classification, quantities and release mechanisms of organic


rhizodeposition (Neumann, 2007).

Environmental influences comprise stress factors with impact on membrane


integrity, such as nutrient limitations (particularly Ca Zn, Mn and P), oxygen also
soil factors such as soil-microbial activity, soil-mechanical impedance and
adsorption characteristics.

Environmental influences comprise stress factors with impact on membrane


integrity, such as nutrient limitations (particularly Ca Zn, Mn and P), oxygen
limitation, temperature extremes, drought and presence of toxic elements, and
also soil factors such as soil-microbial activity, soil-mechanical impedance and
adsorption characteristics.

In some cases these factors exert a more or less unspecific influence on diffusion
mediated release of organic compounds via effects on intracellular concentrations
and leakage of membranes. In other cases they are inductors of highly specific
adaptive responses to counteract environmental stresses, leading to a controlled
release of sometimes enormous amounts of certain root exudates with specific
functions for nutrient acquisition, protection against toxic elements but also as
signals in plant-microbial interactions.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 5

A detailed understanding of the underlying processes is a prerequisite for attempts


to manipulate the quality and quantity of rhizodeposits as a strategy to improve
nutrient acquisition and stress resistance of crops and it may also offer options to
influence the efficiency of microbial fuel cells.

The present contribution tries to give a brief overview, covering the current
knowledge on rhizodeposition and perspectives for a directed manipulation for
practical applications

REFERENCES

Neumann, G. (2007): Root exudates and nutrient cycling. In:Soil Biology Vol. 10.
Marschner P., Rengel Z. (eds.) Nutrient cycling in Ecosystems. Springer, Berlin
Heidelberg, pp.123-157.

Neumann, G., and Römheld, V. (2007): The release of root exudates as affected by
the plant physiological status. In: Pinton, R., Varanini, Z., Nannipieri, Z. (eds.) The
Rhizosphere: Biochemistry and organic substances at the soil-plant interface. 2nd
ed. CRC Press, pp.23- 72.

Neumann, G., Martinoia, E. (2002): Cluster roots - an underground adaptation for


survival in extreme environments. Trends in Plant Science 7 (4), 162-167.

Neumann, G. Römheld, V. (2002): Root-induced changes in the availability of


nutrients in the rhizosphere. In:Plant Roots The Hidden Half, 3rd ed. Waisel Y.,
Eshel, A., Kafkafi U. eds.pp.617-649, Marcel Dekker, New York.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 6
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 7

EXPLORATION OF KEY RHIZOSPHERE PARAMETERS IN


PLANT-MFCS
STEPHAN BLOSSFELD, SVEN SUESSMILCH, CHANTAL A. LE MARIÉ, ARND J.
KUHN

Forschungszentrum Jülich GmbH, IBG-2 Plant Sciences, Germany


Corresponding author E-mail: s.blossfeld@fz-juelich.de

INTRODUCTION

The success of Plant-MFCs is strongly depending on their long-term performance.


Hence, the prerequisite is a stable and long-term power production. However, the
cell potential of plant-MFCs is often not stable of time. For example figure 1
demonstrates the diurnal courses of the cell potentials of two plant-MFCs planted
with Glyceria maxima and Phalaris arundinacea. In both Plant-MFCs the cell
potential fluctuates strongly from night to day. Especially in the case of Phalaris
arundinacea the cell potential shows a diffuse diurnal pattern, whereas Glyceria
maxima shows a clear pattern. There can be several reasons for this dynamic
effect, e.g. changes in root exudation, oxygen release by roots, competition
between different groups of bacteria, temperature effects, etc.

0.20

0.18

0.16

0.14
cell potential (V)

0.12

0.10

0.08

0.06

0.04

0.02

0.00
0:00 8:00 16:00 0:00 8:00 16:00 0:00 8:00 16:00 0:00 8:00 16:00 0:00

Figure 1. In-situ measurements of cell potential of MFC planted with Phalaris


arundinacea (dark line) and Glyceria maxima (light line). The external resistance of
both plant-MFCs was 1000 ohm.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 8

DYNAMICS OF RHIZOSPHERE PARAMETERS

To elucidate these dynamics of MFC cell potential, the non-invasive planar optode
technology was used (Blossfeld and Gansert 2007; Gansert and Blossfeld 2008). By
this, anode internal O2 concentration, pH and CO2 concentration were measured in
the case of Phalaris arundinacea Plant-MFC. The gained information about the pH,
O2 and CO2 dynamics within the MFC will help to improve the understanding of the
short term dynamics of the involved bioprocesses. This is especially necessary for
following the dynamic reactions of the root-rhizosphere processes during
experimental manipulations of the MFC (e.g. change of temperature, light intensity
or nutrient supply). The measurements revealed that the pH dynamics (between pH
6.4 and pH 6.6) in the anode compartment was mainly driven by extremely high
CO2 concentrations in the soil solution of up to 45% during daytime and about 35%
during nighttimes (Fig. 2). Interestingly, the dynamics of the CO2 concentration
does not fit to the course of the cell potential but only to the course of the
temperature (i.e. 18 °C during nighttimes and 24 °C during daytime), hence
especially during daytime, other bacteria seems to dominate in the MFC. The
anode internal oxygen concentration was in a negligible range of less than 1% air
saturation. Hence, in this particular case the diurnal changes of MFC cell potential
are not caused by oxygen release from plant roots.

50 A

45

40

35
p CO2 (%) T(°C)

30

25

20

15

10

0
0:00 8:00 16:00 0:00 8:00 16:00 0:00 8:00 16:00 0:00 8:00 16:00 0:00
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 9

1.0 B 7.0

0.9 6.9

0.8 6.8

0.7 6.7
pO2 (% air saturation)

0.6 6.6

pH
0.5 6.5

0.4 6.4

0.3 6.3

0.2 6.2

0.1 6.1

0.0 6.0
0:00 8:00 16:00 0:00 8:00 16:00 0:00 8:00 16:00 0:00 8:00 16:00 0:00

Figure 2. Non-invasive in-situ measurements of key rhizosphere parameters in a


MFC planted with Phalaris arundinacea. A) CO2 concentration (dark line),
Temperature (light line). B) O2 concentration (dark line), pH (light line).

PERSPECTIVES

Plant-MFCs represent a highly complex system of rhizosphere interactions and


several parameters influence their performance. In this particular case anode
temperature seems to be the key driver of the bioprocesses. Further experiments
will be conducted in order to use this effect for stimulating power production in
Plant-MFCs.

REFERENCES

Blossfeld S and Gansert D 2007 A novel non-invasive optical method for quantitative
visualization of pH dynamics in the rhizosphere of plants. Plant, Cell and
Environment 30, 176-186.

Gansert D and Blossfeld S 2008 The Application of Novel Optical Sensors (Optodes)
in Experimental Plant Ecology. In Progress in Botany. pp 333-358
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 10
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 11

NON-INVASIVE INVESTIGATION OF ROOT GROWTH VIA


NMR IMAGING
STEPHAN BLOSSFELD, CHANTAL A. LE MARIÉ, DAGMAR VAN DUSSCHOTEN,
SVEN SUESSMILCH, ARND J. KUHN

Forschungszentrum Jülich GmbH, IBG-2 Plant Sciences, Germany


Corresponding author E-mail: s.blossfeld@fz-juelich.de

INTRODUCTION

For linking together structure and functioning of plants under varying MFC
conditions, non-invasive quantitative analysis of root growth is necessary. NMR
imaging provides the unique opportunity for a detailed in-situ high resolution 3D
quantification and visualization of the developing root system of individual plants.
For example, NMR imaging can be used for a quantitative comparison of the
development of individual root systems during different environmental treatments,
e.g. changed nutrient supply or water level, over long periods of time. This kind of
structure analysis will help to identify the optimal root system for well performing
MFC.

METHODS & RESULTS

In brief, NMR imaging was done in the EcoNMR facility of the IBG-2 Plant Sciences
Institute (Forschungszentrum Jülich, Germany, www.econmr.org, accessed 18
January 2011) in a 4.7 T Varian VNMRS vertical wide-bore MRI system (Varian Inc.,
Palo Alto, CA,USA) with a quadrature transmit/receive coil and a 300mTm–1
gradient system. Images were obtained by a multi-slice (multi) spin echo technique
with sagittal or axial orientation. For further details we refer to Jahnke et al.
(2009) and Nagel et al. (2009).

First preliminary tests with NMR were done with Glyceria maxima growing in
graphite beads, which is the standard substrate for microbial fuel cells. However, it
turned out that NMR measurements were not possible with graphite as substrate. It
was not possible to receive any usable signal from these measurements (Fig. 1).

The physical reasons for this unexpected reaction of the graphite material are yet
unknown. This result highlights the importance of the substrate characteristics for
NMR studies. High iron or water content can cause severe interferences in the
magnetic fields and therefore disturb the NMR measurements. In this particular
case, graphite needs to be replaced by sand as an alternative substrate in order to
achieve any usable signals (Fig. 2).
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 12

Figure 1. NMR signal (left image) and digital photograph (right image) from a four
week old Glyceria maxima growing in graphite.

Figure 2. NMR image of the root system of a four weeks old Glyceria maxima plant
(left image) and of the same individual a eight weeks later (right image).
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 13

PERSPECTIVES

In a recent experiment, the influence of phosphate deficiency and aluminium


toxicity on root growth was quantified via NMR imaging. This quantitative data is
presented on the 1st international PlantPower symposium (February 10th 2011 in
Gent, Belgium) and will be published soon.

REFERENCES

Jahnke, S., Menzel, M.I., Van Dusschoten, D., Roeb, G.W., Bühler, J., Minwuyelet,
S., Blümler, P., Temperton, V.M., Hombach, T., Streun, M., Beer, S., Khodaverdi,
M., Ziemons, K., Coenen, H.H., Schurr, U., 2009.
Combined MRI–PET dissects dynamic changes in plant structures and functions. The
Plant Journal 59, 634-644.

Nagel, K.A., Kastenholz, B., Jahnke, S., Dusschoten, D.v., Aach, T., Mühlich, M.,
Truhn, D., Scharr, H., Terjung, S., Walter, A., Schurr, U., 2009.
Temperature responses of roots: impact on growth, root system architecture and
implications for phenotyping. Functional Plant Biology 36, 947-959.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 14
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 15

QUANTIFICATION OF EXUDATION FOR THE PLANT-


MICROBIAL FUEL CELL
RENÉ C.P. KUIJKEN1, JAN F.H. SNEL1, HARRO J. BOUWMEESTER2, LEO F.M.
MARCELIS1

1. Wageningen UR Greenhouse Horticulture, Droevendaalsesteeg 1, 6708 PB, Wageningen


2. Wageningen UR Laboratory of Plant Physiology, Droevendaalsesteeg 1, 6708 PB,
Wageningen
Corresponding author E-mail: rene.kuijken@wur.nl

INTRODUCTION & OBJECTIVES

The efficiency and scale of Plant Microbial Fuel Cells (Plant-MFC) need to be
optimized. The amount of
exudates released by the plant is one of the limiting processes of the plant-MFC.
Three strategies to study exudation have been reported: sampling of a non-sterile
root environment (1, 2, 3, 4), sampling of a root environment treated with anti-
microbial agents (5, 6, 7, 8) and sampling of a sterile root environment (9, 10, 11,
12). Initial measurements with HPLC in the non-sterile root zone of tomato plants
showed that exudate concentrations were close to the detection limit. These low
amounts of exudates measured can be the effect of microbial breakdown at the
root-soil interface. Using flow cytometry, we showed that addition of antimicrobial
agents prior to sampling did not kill all bacteria (Kuijken et al. 2010, unpublished).
Net exudation is what drives the energy conversion in the Plant-MFC. The objective
of this study is to develop a simple procedure to estimate net root exudation by
quantifying concentration and breakdown of common exudates in our hydroponic
root system.

MATERIALS & METHODS

Seeds of tomato (Solanum lycopersicon cv Moneymaker, S. lycopersicon cv


Plaisance and S.cheesmanii) were surface sterilized by incubating in 96% EtOH for
15 minutes and in normal commercial bleach with a drop of tween for 20 minutes.
Seeds were 10 times rinsed with sterile deionised water and then put into
antibacterial (600mg/l penicillin + 250mg/l streptomycin) and antifungal (100mg/l
Cycloheximid) solutions for 30 minutes. Seeds were immediately placed on 0.22
(w/v) MS-medium with 0.7% (w/v) of agar in an eppendorf tube with its bottom cut
off. When the sterile plants had developed their first true leaf, they were
transferred to sterile 180 ml tubes in a laminar flow cabinet. Sterile nutrient
solution was added in the flow cabinet. The space between stem and the edge of
the tube was filled with dry cotton balls and covered with leucopore tape. Sterility
of the root system was checked by placing a couple of roots in petri dishes and
detection of contamination on low salt LB medium. Plants were grown in a growth
cabinet at 350 µmol m-2 s-1 (PAR) at 20 °C day and 18 °C night temperature with a
day length of 16 hours and a relative humidity of 80%. Breakdown of exudates was
measured by applying 100 ml of 300 µM artificial acetate solution to the nutrient
medium in the tube and measuring the concentration over time. Average tomato
root dry weight was 130 mg. Organic acids were quantified with a Dionex ICS2500
HPLC system as described (13). Peaks were identified and quantified by co-elution
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 16

of standards with known concentration and the peak surfaces were transformed to
milligrams of anion per liter of exudate solution. With the used HPLC protocol it
was impossible to distinguish between succinate and malate.

RESULTS & DISCUSSION

Figure 1 shows that half of the acetate added at t=0 disappeared from the solution
within 7 hours for cultivars Moneymaker and Plaisance and within 11 hours for S.
cheesmanii. In test systems with a higher root mass/solution volume ratio, half-life
even went down to three hours (data not shown). This can be caused by microbial
metabolism in the biofilm or by uptake by the roots.

250
Cheesmanii Plaisance Moneymaker
Concentration (mg.L .g )
-1

200
-1

150

100

50

0
0 5 10 15 20 25 30
Time (h)

Figure 1. Time course of externally applied acetate in the root environments of


three tomato cultivars. Concentrations are presented in mg L-1 g-1 (root DW). All
starting concentrations at time point 0h were similar at 300 µM.

Figure 2 shows that root exudation leads to an increase in the concentrations of


oxalate (A) and citrate (B) over the 24h observation period. Normalised to root
DW, S. cheesmanii exudes significantly more oxalate, citrate (Fig. 2) and
succinate/malate (not shown) than the other two cultivars. The exudate
concentrations seem to reach a maximum after 12h. At the end of the experiment,
after the samples were taken from the nutrient solution, all roots showed infection
by a monoculture of bacteria. As the roots were sterile prior to the start of the
incubation period (t=0h), these infections must have occurred during or right
before the incubation period. The conditions during incubation can thus be
considered semi-sterile. Experiments to distinguish between uptake of exudates by
the root and by the microbes are under way.

Figure 1 shows that the metabolically active biofilm around the roots has a
significant effect on the amount of acetate in the root zone over time. There is no
reason to assume that the effect of the biofilm on metabolites exuded by the roots
will be much different. This means that for accurate determination of net root
exudation and genetic variation in exudation, without the interfering effect of
microbial activity in the biofilm, the use of a sterile system is inevitable. Current
methods to grow plants with a sterile root zone are not suitable to grow plants to
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 17

beyond the seedling phase or for high-throughput phenotyping for exudation. The
approach presented in this paper requires little effort for plant cultivation and is
suitable for generating the large number of plants required for studying genetic
and environmental effects on root exudation.
12
Cheesmanii Plaisance Moneymaker A
Concentration (mg.L .g )
-1

9
-1

0
0 5 10 15 20 25 30
Time (h)

6
Cheesmanii Plaisance Moneymaker B
Concentration (mg.L .g )
-1
-1

0
0 5 10 15 20 25 30
Time (h)

Figure 2. Time-course of oxalate (A) and citrate (B) in the root environments of
three tomato cultivars. The amount of anions at the different time points is
presented as the concentration in mg L-1 g-1 (root DW).
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 18

REFERENCES

Hoffland E, Vandenboogaard R, Nelemans J, Findenegg G (1992) Biosynthesis and


root exudation of citric and malic-acids in phosphate-starved rape plants. New
Phytologist 122: 675-680

Neumann G, Massonneau A, Martinoia E, Romheld V (1999) Physiological


adaptations to phosphorus deficiency during proteoid root development in white
lupin. Planta 208: 373-382

Hoffland E, Wei CZ, Wissuwa M (2006) Organic anion exudation by lowland rice
(Oryza sativa L.) at zinc and phosphorus deficiency. Plant and Soil 283: 155-162

Pearse SJ, Veneklaas EJ, Cawthray G, Bolland MDA, Lambers H (2007) Carboxylate
composition of root exudates does not relate consistently to a crop species' ability
to use phosphorus from aluminium, iron or calcium phosphate sources. New
Phytologist 173: 181-190

Imas P, BarYosef B, Kafkafi U, GanmoreNeumann R (1997) Phosphate induced


carboxylate and proton release by tomato roots. Plant and Soil 191: 35-39

Imas P, BarYosef B, Kafkafi U, GanmoreNeumann R (1997) Release of carboxylic


anions and protons by tomato roots in response to ammonium nitrate ratio and pH
in nutrient solution. Plant and Soil 191: 27-34

Gherardi MJ, Rengel Z (2004) The effect of manganese supply on exudation of


carboxylates by roots of lucerne (Medicago sativa). Plant and Soil 260: 271-282

Gent MPN, Parrish ZD, White JC (2005) Nutrient uptake among subspecies of
Cucurbita pepo L. is related to exudation of citric acid. Journal of the American
Society for Horticultural Science 130: 782-788

Meharg AA, Killham K (1991) A novel method of quantifying root exudation in the
presence of soil microflora. Plant and Soil 133: 111-116

Gaume A, Machler F, De Leon C, Narro L, Frossard E (2001) Low-P tolerance by


maize (Zea mays L.) genotypes: Significance of root growth, and organic acids and
acid phosphatase root exudation. Plant and Soil 228: 253-26411

Sandnes A, Eldhuset TD, Wollebaek G (2005) Organic acids in root exudates and soil
solution of Norway spruce and silver birch. Soil Biology & Biochemistry 37: 259-269

Kamilova F, Kravchenko LV, Shaposhnikov AI, Azarova T, Makarova N, Lugtenberg B


(2006) Organic Acids, Sugars, and l-Tryptophane in Exudates of Vegetables Growing
on Stonewool and Their Effects on Activities of Rhizosphere Bacteria. Molecular
Plant-Microbe Interactions 19: 250-256

Bentsink L, Yuan K, Koornneef M, Vreugdenhil D (2003) The genetics of phytate and


phosphate accumulation in seeds and leaves of Arabidopsis thaliana, using natural
variation. Theoretical and Applied Genetics 106: 1234-1243
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 19

NUTRIENT SUPPLY INFLUENCES ROOT EXUDATION


AND DEVELOPMENT
CHANTAL LE MARIÉ, STEPHAN BLOSSFELD, SVEN SÜßMILCH, ARND J.
KUHN

Research Center Jülich, IBG-2 Plant Sciences, Germany


Corresponding author E-mail: s.blossfeld@fz-juelich.de

INTRODUCTION

The objective of this study was to investigate the effect of the composition of the
nutrient solution on the exudation of various plant species under waterlogged
conditions. Root exudation is influenced by the root environment, especially by the
availability of nutrients in the soil or ion toxicity. It is known that phosphate
deficiency as well as aluminum addition can increase the exudation of
carbohydrates such as citrate and malate (Delhaize 2001; Hoffland et al. 2006;
Wenzl et al. 2002). Thus we tried to increase the exudation of several plant species
by treating them with phosphate deficiency or aluminum addition under
waterlogged conditions. Furthermore the influence of the treatments and the
waterlogged conditions on the root development and plant health was studied.

METHODS AND RESULTS

During the first experiments a Hoagland solution (diluted 1:1 with water) and two
modified Hoagland nutrient solutions (diluted 1:1 with water before modified) were
used: a) Hoagland + 2 mM Aluminum chloride (AlCl3); b) Hoagland containing only
10% of normal phosphate (PO43-). In a later stage of the experiments, solution a)
contained only 1mM AlCl3. The reason for this change was the dramatic effect of
the treatment on plant health like root rot or reduced growth (see Figure 1). In
later experiments with reduced AlCl3 content no dramatic changes in root health
and biomass were detectable (see Figure 2).

These nutrient solutions were applied to various plant species (Phalaris


arundinacea, Glyceria maxima, Lythrum salicaria, Arundinella anomala, Spartina
anglica, Hemathria spec. and Oryza sativa) and soil solution samples were taken at
different time intervals. The samples were analyzed via capillary electrophoresis
analysis (CE), enzymatic assays and total organic carbon (TOC) analysis in order to
quantify the influence of the changed nutrient solution on root exudation.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 20

Hoagland control Phosphate deficiency Aluminum addition

Figure 1. Comparison of the effect of the different Hoagland solutions on the


growth of Glyceria maxima, showing a strong effect after aluminum addition on
root growth and health.

Total organic carbon measurements indicated that phosphate deficiency as well as


aluminum treatment influences exudation. Especially the total organic carbon
content was enhanced by phosphate deficiency 72h after starting the treatment.
However for aluminum was no significant increase of TOC content measurable after
72 h. On the contrary, in short-term experiment with a high number of samples
within 12 hours after treatment, an increase of the TOC content was observable
already 1.5 h after aluminum addition. This indicates that the reaction on
aluminum is really fast and occurs immediately after the treatment.

Detectable by capillary electrophoresis were different compositions and


concentrations of the org. acids depending on the chosen treatment (phosphate
deficiency or aluminum addition) but the sum of the organic acid content remained
constant.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 21

Hoagland control Aluminium addition

Figure 2. Root development of Glyceria maxima after treatment with Hoagland


containing 1mM AlCl3.

OUTLOOK

The future purpose will be to check if a treatment with phosphate deficiency or


aluminum addition is a possible tool to increase the productivity of the microbial
fuel cell without affecting the plant development and health.

REFERENCES

Delhaize E 2001 The role of root exudates in aluminum tolerance. Springer-Verlag


Tokyo, Tokyo. pp. 140-155.

Hoffland E, Wei C Z and Wissuwa M 2006 Organic anion exudation by lowland rice
(Oryza sativa L.) at zinc and phosphorus deficiency. Plant and Soil 283, 155-162.

Wenzl P, Chaves A L, Patino G M, Mayer J E and Rao I M 2002 Aluminum stress


stimulates the accumulation of organic acids in root apices of Brachiaria species.
Journal of Plant Nutrition and Soil Science 165, 582-588.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 22
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 23

B:
MICROBIAL ASPECTS OF THE
RHIZOSPHERE & ANODE
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 24

KEYNOTE BY PROF. DR. MICHAEL W. FRIEDRICH

The Microbial Ecophysiology lab has been founded in June 2009 at the
Faculty of Biology/Chemistry, University of Bremen, Germany. The major
focus of the group is the to link structure ( “who is there”) to function
(”what are they doing”) of microbial communities. We are interested in
structure-function relationships of anaerobes (1) that respire anaerobically
(e.g., dissimilatory iron reducers, methanogens, dehalogenating
microorganisms, electrigens), and (2) play an important role in the carbon
flow through microbial communities in a number of environments such as
rice paddy soil, fresh water, and marine sediments. Central to our
approach is the state-of the-art community level analyses with molecular
tools (e.g., stable isotope probing) in combination with an assessment of
the underlying biogeochemical processes.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 25

THE MICROBIAL ECOLOGY OF ELECTRIGENIC


MICROORGANISMS IN PLANT-RHIZOSPHERE BASED
MICROBIAL FUEL CELLS

MICHAEL W. FRIEDRICH

Microbial Ecophysiology, Faculty of Biology/Chemistry, University of Bremen, Germany


Corresponding author E-mail: michael.friedrich@uni-bremen.de

Microbial fuel cells (MFC) are electrochemical devices in which microorganisms are
involved in the generation of electrical current. Certain microbes are capable of
transferring electrons from the oxidation of a substrate to an anode, as well as
receiving electrons at the cathode (biocathode). A rather novel development in the
field is the plant-rhizosphere based microbial fuel cell (1,2,3). These systems
capitalize on the photosynthetically fixed carbon that is naturally supplied to the
rhizospheric microbial community of wetland plants such as rice. With anodes
buried in the rhizosphere, ultimately becoming overgrown and even penetrated by
plant roots, anode affiliated microorganisms can directly tap into the flow of
exudates from the plant, oxidize the photosynthesates, and transfer electrons to
the anode, thereby generating current in closed circuit systems. Compared to
canonical sediment fuel cells, an increase in power production of up to 7-fold has
been observed in rice-based MFC (1), and current densities of up to 67 mW m-2
total anode surface area have been attained (3). Naturally, rhizodeposits, which
are products of photosynthesis and thereby of the unlimited energy source light,
will be microbially converted to the greenhouse gas methane; therefore, it is a
remarkable opportunity to expand the possibilities for green-energy generation by
plant-based MFC technology.

Since microorganisms are a key component for the current generation in these
systems, one of the first steps has been to assess, which microorganisms are
involved using cultivation independent molecular tools such as 16S rRNA gene based
analyses. The presence of a rice plant as well as the availability of a closed circuit
system ultimately shape the microbial community on the rhizosphere influenced
anodes, pointing clearly to a coupling of rhizodeposit oxidation linked to electron
transfer to the anode (4). Furthermore, the support for rice plants (potting soil vs.
rice field soil) as well as the inoculum (e.g., rice field soil vs. MFC reactor effluent)
is a key determinant of community composition. Potting soil stimulates delta-
proteobacterial Desulfobulbus spp. the most, whereas rice field soil selects for
delta-proteobacterial Geobacter and Anaeromyxobacter spp. on anode surfaces in
closed circuit systems. The presence of these electrigenic microorganisms gives
first clues on the potential mechanisms involved in electron transfer to anodes. In
case of Desulfobulbus spp., known sulfate-reducing microorganisms previously
encountered on marine sediment MFC anodes, it is suggestive to assume a potential
involvement of sulfur-cycling, whereas Geobacter spp., are known iron-reducing
microorganisms in rice field soils (5) and as being capable of transferring electrons
directly to the anode surface. How methanogenic Archaea are affected by the
presence of anodes in rice planted MFCs is not yet understood. Methanogenic
Archaea do occur on anodes. In fact, a closed circuit system favors the abundance
of hydrogenotrophic over acetoclastic methanogenic Archaea and decreases
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 26

acetate availability. Thus, the presence of anodes might increase competition for
the common resource acetate in rice planted MFC microcosms, which opens vistas
to another application of plant-based MFCs: regulation of methane production. At
the field scale, however, methane emission has been found unaffected (2)
underpinning that future efforts are necessary to understand factors limiting the
efficiency of plant-based MFCs.

REFERENCES

1 DeSchamphelaire et al. (2008) Environ. Sci. Technol.

2 Kaku et al. (2008) Appl. Microbiol. Biotechnol.

3 Strik et al. (2008) Int. J. Ener. Res.

4 DeSchamphelaire et al. (2010) Appl. Environ. Microbiol.

5 Hori et al. (2010) ISME J


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 27

MICROBIAL RESOURCE MANAGEMENT (MRM): THEORY


AND PRACTICAL TOOLS TO EXPLOIT BACTERIAL
CAPABILITIES

MASSIMO MARZORATI, SUZANNE READ, JAN B.A. ARENDS, WILLY


VERSTRAETE, NICO BOON

LabMET, Ghent University, Coupure links 653, 9000 Gent Belgium.


Corresponding author E-mail: massimo.marzorati@ugent.be

INTRODUCTION

Microbes represent nearly half of all the biomass on earth and catalyze almost all
the biogeochemical cycles in the biosphere. This huge metabolic reservoir is an
extremely important resource ready to be exploited in order to generate new
products and processes, improve human and environmental health and assure
environmental sustainability. The exploitation of bacteria to try to solve practical
problems has been defined as Microbial Resource Management similarly to the well-
known concept of Human Resource Management (Verstraete et al., 2007).
The first step, in order to harness the unique abilities associated to microbial
communities, is the understanding of the ecological principles that regulate this
complex interactions. For this purpose, community-level molecular techniques are
widely used in comparative microbial ecology to assess the diversity of microbial
communities and their response to changing environments. However, the amount
of data derived from these techniques is continuously increasing and the lack of a
universal interpretation makes impossible the determination of predictive
behaviours.

THEORY

We have developed a specific tool-set for a setting-independent theoretical


interpretation of the raw fingerprinting data based on three levels of analysis: (i)
the range-weighted richness (Rr) reflecting the carrying capacity of the system, (ii)
the dynamics (Dy) reflecting the specific rate of species coming to significance, and
(iii) the community organization (Co), defined by the structure of a microbial
community in terms of evenness (Marzorati et al., 2008). We also showed that the
latter is a key parameter to preserve the community functionality under selective
stress conditions (Wittebolle et al., 2009). When communities are highly uneven, or
there is extreme dominance by one or a few species, their functioning is less
resistant to environmental stress (Fig. 1).

These three parameters have been used to describe the most disparate
environments but the possibility of steering the capabilities of a microbial
community for practical applications is in many cases a far goal. Further
fundamental research is much needed in this area in order to move a step forward
and go from the ecological description of a given ecosystem to the prediction of its
functionality under changing environmental conditions.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 28
Func onality 

Evenness 
Figure 1. Microbial functionality in relation to the evenness. A selective stressor
has a much more negative impact on the functionality of the unevenly distributed
microbial community.

EXAMPLE

As an example, we calculated the Co parameter for the bacterial and archaeal


communities (T-RFLP analyses) residing on the anodes of microbial fuel cells
inserted in the rhizosphere of living rice plants (Tab. 1) (De Schamphelaire et al.,
2010).

Table 1. Co parameter calculated for the bacterial and archaeal communities of


the anode in presence of absence of plants (CC = closed circuit).
Com m unit y or ga niza t ion
Ba ct Pla nt CC 59.7
Ba ct N o Pla n t CC 45.8

Arch Pla nt CC 53.2


Arch N o Pla n t CC 42.5

The main finding of the paper was that both microbial communities were
influenced by the excreted organic compounds, the sediment matrix and the
electrical connection (De Schamphelaire et al., 2010). Co values indicated that the
presence of plants induced a selection for some species (i.e. Desulfobulbus-like
spp.) in the bacterial community thus leading to a more uneven structure. The
same conditions moved the Archaeal community in the opposite direction, towards
a more even community. The strictly acetotrophic Methanosaetaceae decreased
with a concomitant increase of Methanobacterium and Methanosarcina, possibly
leading to a more hydrogenotrophic methanogenesis.

PERSPECTIVE

MRM can thus be a useful tool in monitoring reactor performance. It can also be
used to improve or steer underperforming biological reactors. The Plant powered
MFC can also be considered a reactor where the stability and functionality of the
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 29

microbial community performing a key task can be monitored and possibly steered
by means of MRM. The microbial community is not only key to power production
but also indispensible for plant health. Therefore close monitoring and possibly
steering of the microbial community can greatly enhance the understanding and
output of complex biological reactors such as the Plant powered fuel cell.

REFERENCES

De Schamphelaire L, et al. (2010) Microbial community analysis of anodes from


sediment microbial fuel cells powered by rhizodeposits of living rice plants. Appl
Environ Microbiol. 76:2002-2008

Marzorati M, et al. (2008) How to get more out of molecular fingerprints: practical
tools for microbial ecology. Environ Microbiol 10:1571-1581

Verstraete W, et al. (2007) Microbial resource management: The road to go for


environmental biotechnology. Eng Life Sci 7:117-126

Wittebolle L, et al. (2009a) Initial community evenness favours functionality under


selective stress. Nature 458:623-626
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 30
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 31

COMPARISON OF BACTERIAL RHIZOSPHERE


COMMUNITIES FROM PLANT MICROBIAL FUEL CELLS
WITH DIFFERENT CURRENT PRODUCTION BY 454
AMPLICON SEQUENCING
M. ROTHBALLER1, M. ENGEL2, D.P.T.B.T. STRIK3, R. TIMMERS3, M.
SCHLOTER2, A. HARTMANN1
1
Department Microbe-Plant Interactions,
2
Institute of Soil Ecology/Department Terrestrial Ecogenetics, Helmholtz Zentrum München,
German Research Center for Environmental Health, Ingolstädter Landstraße1, 85764
Neuherberg,
3
Sub-department of Environmental Technology, Wageningen University, 6703 HD Wageningen,
The Netherlands
Corresponding author E-mail: rothballer@helmholtz-muenchen.de

With Glyceria maxima root samples, which were derived from the anode
compartments of one plant MFC which produced high current output and one with
lower current, a 454 pyrosequencing run was performed. From each of these MFCs
root samples from the upper and the lower area of the MFC were taken. After DNA
extraction and PCR amplification using a pair of 16S rDNA targeted universal
bacterial primers meeting the requirements of the 454 sequencing protocol, the
PCR product was purified by gel extraction. Two different annealing temperatures
were used (50°C and 54°C) which resulted in a total of 8 samples. The different
samples were tagged with unique multiplex identifier sequences (MIDs), which are
detectable by the sequencing software tool. Therefore all samples could be pooled
and sequenced on one quarter of a pico titer plate of the Roche GS FLX Titanium
sequencing platform.

In most cases an annealing temperature of 50°C resulted in a higher diversity and


total number of sequences. There were no major qualitative differences between
the two annealing temperatures. These first results provided already a good
overview of the microbiological biodiversity in the two systems but the sizes of the
sequenced fragments were not in all cases optimal. This was probably due to the
purification of the amplicons by gel extraction, which to some extent also produces
smaller DNA fragments that are preferably sequenced by the 454 system. Therefore
the sequencing run was repeated after an additional purification step with AMPure
beads (Agencourt, Beckman Coulter), which bind only fragments larger than 150
bp. With this method it was possible to receive optimal sequencing results with
readlengths at an average of about 500 bp. These sequences were then grouped
into the different samples with the help of the MIDs and could be assembled by the
Newbler software (Roche) with a similarity value of 99% and an overlap of 400 bp.
The resulting consensus sequences were imported into the ARB phylogenetic
software tool, 16S sequences were inspected for chimera by the Bellerophon
software tool (http://foo.maths.uq.edu.au/ _huber/bellerophon.pl), aligned
automatically, corrected manually and allocated to their proper position in the
phylogentic tree.

Microbiological diversity was found to be highest for the high current producing
plant MFC roots from the top layer, while microbiological differences between high
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 32

and low current producing plant MFCs were most pronounced for roots from the
bottom layer. Most dominant classes were Clostridia in the high current producing
plant MFCs and β-Proteobacteria in the low current system. The families of
Ruminococcaceae for the high current plant MFC and Comamonadaceae for the low
current producing plant MFC were the dominant groups, making up over 50% of the
total 16S rDNA sequences found in the samples from the bottom part of the plant
MFCs. For details see table 1.

To localize active bacteria detected with 454 sequencing of the 16S rDNA, the
fluorescent in situ hybridization (FISH) technique was applied. With a Geobacter
specific probe set it was possible to detect fluorescent Geobacter cells which
appeared more frequently on graphite granules than root surfaces. With a probe
specific for only three Ruminococcaceae species a small number of cells was
positively identified on the surface but to a much larger extent fluorescent signals
were detected in outer cortex layers of roots of the high current plant MFC.

Table 1. Characteristic families for the different samples. Families were defined as
characteristic for one sample, if the number of sequences found in that single
sample was over 40 % of the total amount of sequences allocated to that family in
all 4 samples together.

High current Low current


top Anaerolineae, Caldilineae, Propionibacteraceae,
Eubacteriaceae, Haliangiaceae
Planctomycetaceae,
Hyphomicrobiaceae,
Methylocystaceae, Rhizobiaceae,
Xanthobacteraceae, Opitutaceae,
Veillonellaceae, Holophagaceae
bottom Geobacteraceae, Veillonellaceae, Prevotellaceae, Neisseriaceae,
Ruminococcaceae, Clostridiaceae, Oxalobacteraceae,
Bacteroidaceae, Comamonadaceae,
Acidobacteriaceae Cystobacterineae,
Spirochaetaceae

Archaeal 16S rDNA was more difficult to amplify from the extracted DNA than
bacterial 16S, which accounts for a low DNA content compared to the bacteria.
There was a three times higher amplificate concentration detectable from samples
derived from low current producing plant MFCs compared to the high current
system, indicating that Archaea in total were more abundant in the low current
plant MFC. Diversity of Archaea was much less then of Bacteria. In the low current
plant MFC members of the Methanobacteriaceae family were accompanied by
several genera belonging to other families, while in the high current plant MFC the
genus Methanobacterium was making up over 95% of the total community.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 33

METHANOTROPHIC MICROBIOME
INKA VANWONTERGHEM, SUZANNE READ, DAVID VAN DER HA, WILLY
VERSTRAETE, NICO BOON

Ghent University, Laboratory of Microbial Ecology and Technology (LabMET)


Coupure 653, 9000 Ghent, Belgium
Corresponding author E-mail: Nico.boon@ugent.be

Introduction and objectives


Due to our increasing global warming issues, current research is focusing on new
techniques to enhance biological methane mitigation. One such technique is the
use of microbial fuel cells (MFCs) in rice paddies which are a prominent
anthropogenic methane source. It is hypothesized that microbial catalyzed
oxidation of organic material in MFCs diminishes the amount of substrate left for
methanogenesis, thus decreasing methane production and subsequent emission [1].
Another option is to stimulate methane oxidizing bacteria (MOBs) in the rice plant
rhizosphere, thereby increasing methane oxidation. This research project
investigates specific interactions between methanotrophic and heterotrophic
bacteria in order to attain higher methane oxidation activity. It is well-known that
MOBs grow better and show higher activity in mixed cultures compared to pure
cultures. Thus, it is hypothesized that this higher oxidation rate is either due to
removal of possibly toxic intermediates or the production of growth factors [2-5]. A
recent discovery has also suggested that bacteriocins, produced by Enterococcus
faecium, increase methanotrophic activity through a possible stress-related
response of the MOBs. The aim of this project is to investigate these specific
interactions in order to stimulate methanotrophic communities in soils to increase
methane oxidation, and thus decrease methane emissions to the atmosphere. To
achieve this we used cocultures of MOBs and heterotrophs which have been well
characterized and have previously been identifies in stable methanotrophic-
heterotrophic communities [2-4,6]. This application can also be performed using
MFCs as a platform to test the decrease of methanogenic methane production with
the efficiency of methanotrophic methane oxidation.

Materials and methods

Pure culture growth


Selected MOBs were Methylococcus capsulatus (M. capsulatus) (NCIMB11853T),
Methylocystis hirsuta (M. hirsuta) (DSM18500T), Methylocystis parvus (M. parvus)
(NCIMB11129T) and Methylosinus trichosporium (M. trichosporium) (NCIMB11131T).
MOBs were grown in dilute Mineral Salts Medium (dNMS) [7], at pH 7 and 20 vol%
methane was added. M. capsulatus was grown at 37°C, the others at 28°C, and all
cultures were placed on a shaker at 120 rpm. Closed Schott bottles, with a volume
of 100 mL were used (Schott AG, Mainz, Germany) and sealed with a rubber butyl
stopper (Ochs GmbH, Bovenden, Germany). Heterotrophs were grown in Luria-
Bertani medium (1% tryptone, 0.5% yeast extract and 0.5% NaCl), at 28 °C and
placed on a shaker at 120 rmp. An initial broad screening test in 96-well plates led
to the selection of the following heterotrophic bacteria: Pseudomonas aeruginosa
(P. aeruginosa) (LMG1242), Pseudomonas stutzeri (P. stutzeri) (LMG1228),
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 34

Rhodobacter sphaeroides (R. sphaeroides) (LMG2823), Blastobacter denitrificans


(B. denitrificans) (LMG8443), Shewanella oneidensis (S. oneidensis) (LMG19005),
Enterococcus faecium (E. faecium) (LMG8147), Xanthomonas autotrophicus (X.
autotrophicus) (LMG7043) and isolate Nocardioides sp.

Co-culture growth
MOBs were grown for 3 days before adding the heterotrophs at cell densities of
1000 lower than the MOBs determined using flow cytometry. The heterotrophs
were washed twice with dNMS before being added to the pure methanotroph
culture. Co-cultures were grown in the same conditions as pure methanotrophic
cultures and all experiments were performed in triplicates.

Activity measurements
GC-measurements were taken to determine oxygen, methane and carbon dioxide
content. Pressure measurements were also taken to determine the final % of
methane oxidation.

Results and discussion

Addition of heterotrophic bacteria significantly increased the methane oxidation


capacity of the methanotrophs (Figure 1). When grown in co-culture with P.
aeruginosa the methane oxidation activity of M. capsulatus increased, leading to a
decrease in methane content of 60.5 ± 4.5 % within 10 days, this compared to a
decrease of 20.2 ± 4.7 % when M. capsulatus was grown in pure culture (Figure 1A).
Co-culturing with E. faecium led to a decrease in methane content of 63.8 ± 1.6 %
(Figure 1A). Another experiment performed with M. hirsuta indicated an increase
in methane oxidation activity due to the presence of Nocardioides sp. an isolate
found in association with MOBs (unpublished data). When grown in co-culture with
Nocardioides sp. the methane content decreased with 64.8 ± 2.9 %, compared to a
decrease of 34.5 ± 12.5 % when M. hirsuta was grown in pure culture (Figure 1B). In
contrast, the heterotrophic bacteria showed no growth or activity when grown with
methane as sole energy and carbon source. These experiments allow us to conclude
there is an increase in methane oxidation due to the presence of certain
heterotrophic bacteria and there appears to be a specific methanotroph-
heterotroph interaction.

Further research to investigate whether the higher oxidation rate is either due to
removal of possibly toxic intermediates or to the production of growth factors is
ongoing. Supernatant experiments will be performed for these purposes, and these
experiments consist of growing heterotrophs, adding the supernatant to the pure
methanotroph cultures and observing methane oxidation activity. An initial
experiment has already been performed, where the supernatant of E. faecium was
added to the pure M. capsulatus culture. The supernatant presumably contained
enterocin, a bacteriocin produced by E. faecium, and addition of the supernatant
led to a decrease in methane content of 63.0 ± 3.7% compared to a decrease of
20.2 ± 4.7% when M. capsulatus was grown in pure culture (unpublished data).
Additionally, growth yield measurements will be performed to investigate the
increase in biomass, and the mechanistic of the microbiome will be explored by
analyzing the composition of the organics generated by the bacteria, organics such
as PHB, EPS and proteins.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 35

1
0,9
0,8
0,7
CCH4 / C0,CH4

0,6
0,5
0,4
0,3
0,2
A
0,1
0
0 50 100 150 200 250
Time after innoculation (hrs)

1
0,9
0,8
0,7
CCH4 / C0,CH4

0,6
0,5
0,4
0,3
0,2
0,1 B
0
0 50 100 150 200 250
Time after innoculation (hrs)

Figure 1. Methane oxidation activity of strain (A) M. capsulatus (MC) and (B) M.
hirsuta (MH) in pure cultures and in co-cultures. Symbols: (A) pure MC-culture,
co-culture with P. aeruginosa, co-culture with E. faecium;
(B) pure MH-culture, co-culture with Nocardioides sp.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 36

Further in-depth research using Fluorescent In-Situ Hybridization (FISH) [8]


followed by imaging with confocal microscopy will allow for biofilm quantification
and the determination of methanotroph/heterotroph biomass ratios. The presence
of both species close together in a biofilm is also an indication of interaction
between the two species. Stable Isotope Probing (SIP) [9] will be performed to look
at the nutrient cycling and to indicate the presence of a syntrophic interaction
between the methanotrophic and heterotrophic bacteria.

Finally, a comparison can be made between the effectiveness of the two following
methane mitigation strategies: first, the increased methane oxidation activity due
to a specific methanotrophic-heterotrophic interaction and second, the decreased
methane production via microbial fuel cells.

References

1. De Schamphelaire, L., et al., Microbial fuel cells generating electricity from


rhizodeposits of rice plants. Environmental Science & Technology, 2008. 42(8): p.
3053-3058.

2. Hrsak, D. and A. Begonja, Possible interactions within a methanotrophic-


heterotrophic groundwater community able to transform linear
alkylbenzenesulfonates. Appl Environ Microbiol, 2000. 66(10): p. 4433-4439.

3. Hrsak, D. and A. Begonja, Growth characteristics and metabolic activities of


the methanotrophic-heterotrophic groundwater community. Journal of Applied
Microbiology, 1998. 85: p. 448-456.

4. Hesselsoe, M., et al., Degradation of organic pollutants by methane grown


microbial consortia. Biodegradation, 2005. 16(5): p. 435-448.

5. Jiang, H., et al., Methanotrophs: Multifunctional bacteria with promising


applications in environmental bioengineering. Biochemical Engineering Journal,
2010. 49(3): p. 277-288.

6. van Bodegom, P., et al., Methane oxidation and the competition for oxygen in
the rice rhizosphere. Applied and Environmental Microbiology, 2001. 67(8): p.
3586-3597.

7. Whittenbury, R., K.C. Phillips, and J.F. Wilkinson, Enrichment, isolation and
some properties of methane-utilizing bacteria. J Gen Microbiol, 1970. 61: p. 205-
217.

8. Amann, R.I., W. Ludwig, and K.H. Schleifer, Phylogenetic identification and in


situ detection of individual microbial cells without cultivation. Microbiology
Reviews, 1995. 59: p. 143-169.

9. Neufeld, J.D., et al., DNA stable-isotope probing. Nature protocols, 2007. 2: p.


860-866.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 37

C:
MICROBIAL FUEL CELL
TECHNOLOGY
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 38

KEYNOTE BY PROF. DR. JOHN GREENMAN

BRL is a partnership platform between the University of the west of


England, Bristol and the University of Bristol. The lab’s mission is to
understand the science, engineering and social role of robotics and
embedded intelligence. Our multidisciplinary approach aims to create
autonomous devices capable of working independently, with each other, or
with us in our human society.

For over a decade we have developed a group, within the BRL, that is
particularly interested in energy autonomy for artificial agents. The main
thrust of the research for this group is on the Microbial Fuel Cell
technology, which when configured as a stack it is used to power EcoBot
robots I, II and III, which demonstrate the principle of self-sustainability
i.e. the ability to collect organic fuel (biomass) from the environment,
metabolise it to generate electricity, manage this electrical energy
onboard and excrete the waste. In addition, the group has delved into
wastewater treatment using the MFC stack technology – a project funded
by industry and UK’s Engineering and Physical Sciences Research Council
(EPSRC).

BRL is part of the EPSRC’s SUPERGEN-V Consortium, which focuses research


on biological fuel cells. BRL is also part of another EPSRC’s Grand
Challenge Consortium that is looking into methods of CO2 capture and
utilisation using metal organic frameworks (MOFs) and algae in MFCs.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 39

OVERVIEW OF MICROBIAL FUEL CELLS FOR PRACTICAL


POWER PRODUCTION

GREENMAN J AND IEROPOULOS I

Bristol Robotics Laboratory, UWE, Bristol, BS16 1QY, United Kingdom


Corresponding author E-mail: john.greenman@uwe.ac.uk

Microbial Fuel Cells (MFCs) are bio-electrochemical transducers, which convert


biochemical energy from organic fuels (including low grade wastes such as
anaerobic sludge, landfill-leachate, insects and plant-materials, including
microalgae) into electricity, through the reactions of microbes growing as biofilms
around the electrodes. The earliest description of such a device was that of Potter
(1912) with outputs of a few microWatts per MFC. In contrast, MFC’s described
more recently in the literature are more powerful by a factor of 2-log fold or
greater (Rabaey et al. 2003; Cheng et al. 2006).

However, a single MFC can never produce more than the theoretical maximum of
1.14V (open circuit voltage – VO/C) and typically VO/C values are of the order of 0.7V
which decreases to about half (typically 0.35V) when switched to a load resistor at
maximum power transfer. At this voltage, the power produced is generally
insufficient to energize electronic devices or charge up accumulators. Thus,
pluralities of MFCs (i.e. 2 or more in a stack) are likely requirements for running
any applications.

The first published study of MFC stacks (Cohen, 1931) reported an output of 2mA at
35V (70mW) from a stack of an unspecified number of units (estimated to be
around 70 units). However, the author gave insufficient information regarding the
experimental working conditions and did not comment on the sustainability of
power output [from batch culture systems]. Further developments of MFC stacks in
practical applications were made by Wilkinson (2000), Ieropoulos et al. (2003) and
Ieropoulos et al. (2005). Aelterman et al. (2006) described a stack of MFC
configured to work in continuous fluid flow. The units were connected together
both fluidically and electrically, although details of fluidic isolation, which is
critical for the operation of such stacks (Ieropoulos et al. 2008), were not
presented or discussed. Nevertheless, this study was the first to report cell reversal
in a series-connected stack.

With regard to scale, it seems likely that more efficient energy harvesting takes
place in small-scale MFC units and, thus, there is a natural drive for miniaturisation
and multiple-unit stack development. The latter carries its own challenges, some
of which are associated with internal resistance, cell polarity reversal and electron
leakage across structural materials. One important feature of stacks of MFC
connected electrically and fluidically together is that electrons flow through the
fluidic lines of interlinked MFCs, thus causing fluidic conductance, which opposes
the voltage increases that would be expected from series connected stacks, (i.e.
shunt losses) (Ieropoulos et al. 2008). It therefore becomes vital how such stacks
may be configured electrically and fluidically for high performance and stability
(see figure 1).
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 40

- - +

+
Cathode

Cathode

Cathode

Cathode

Cathode

Cathode
Anode

Anode

Anode

Anode

Anode

Anode
(a) (b)

- +
- +

Cathode
Cathode

Cathode

Cathode
Cathode

Cathode

Cathode

Anode
Anode

Anode

Anode
Anode

Anode

Anode

(c) (d)

Figure 1. Schematic representation of the different fluidic and electrical


configurations for stacks of MFCs: (a) parallel electrical connection with a common
feed line; (b) series electrical connection with a common feed line; (c) series
electrical connection but with individual feed lines; and (d) series–parallel
connections with individual feed lines and with an even number of MFCs.

In essence, MFC units need to be isolated from each other, to prevent any electron
leakage through the various futile paths. The schematic in Fig. 1 illustrates
multiple MFC units connected in a series/parallel mode and fed from individual
feedstock bottles. The latter is simply a representation of the isolation element
that is required and symbolically shows how MFC units can be efficiently
maintained.

Even though stacks of MFCs are more powerful than individual units, they are still
only suitable for powering the lowest power requiring devices. Such devices must
be able to manage the energy – both that produced and spent – by incorporating
high efficiency “electron-harvesting” systems, which extract electrons by adjusting
the load and current output to persistently charge a capacitor (accumulator) at
maximum sustainable power transfer. One example of such a system, is that
devised, built and used to operate EcoBots I and II and control the output from 8
MFCs in order to power the robot in what is called “pulsed behaviour mode”
(Ieropoulos et al. 2003; Melhuish et al. 2006). EcoBot was built around a stack of 48
small-scale MFCs and demonstrated “energy autonomy” i.e. the ability to get food
and water from its environment, to digest the food and circulate the digest through
the onboard stack of MFCs using mechatronic units and to get rid of its own waste.
This required the generation and storage of sufficient energy to run a total of 5
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 41

motors and 4 pumps. For any large number of continuously fed units in stacks,
there is an energy cost associated with the fluid flow of input and output. Although
a gravity gradient may assist, there is a need for at least one reliable feed-pump
and these require considerable energy. A true self-sustainable system must produce
enough energy to run its peripheral maintenance modules. This is not a trivial
challenge but unless it is met, such a system will never be a net energy provider.

Figure 2. Stacks of 48 MFC capable of producing 2mW of power when configured in


series–parallel connections. Inset shows four designs of MFC made from different
polymeric materials; all similar when tested as units, but some unsuitable when
configured in a stack

STACK PROPERTIES
Internal resistance is probably the most critical measurement that is required for
understanding performance of MFCs in stacks. There are several ways of measuring
this parameter and although EIS is well established in the field of electrochemistry,
this method can only accurately calculate the ohmic part of the total internal
losses. Polarisation sweeps on the other hand allow the holistic quantification of
the total internal losses, without selectively identifying which part is contributing
how much to the total internal resistance, and this is a method often preferred by
workers in the field. There are however questions around the sweep time
(resistance connection time), which still need addressing.

Cell reversal
In physics, polarity reversal is a phenomenon often observed in networks (stacks) of
multiple electrical devices – such as batteries – connected together. The reversal
takes place when the units are connected in series, because this is when the
voltage differential pressure is at its highest and, thus, is more sensitive to any
changes in the internal state of each of the interconnected units.

THE BIOTIC COMPONENTS OF MFCs


Complex mixed culture biofilms are invariably formed after inoculation from any
anaerobic sludge or sediment samples. The diverse nature of the anodic biofilms
means that most types of soluble organic carbon (sugars, acetate, lactate, amino
acids) can be utilised by the bioelectrode system. More complex feedstocks
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 42

(macromolecular; sometimes particulate; including microalgae and plant materials)


may require a longer residence time in the stack system to be fully hydrolysed or
“digested”. In highly diverse bacterial communities, there are many species that
can “substitute” for one another with regard to the main functional properties of
the microcosm (i.e hydrolysis/utilisation of carbon-energy substrates and electron
transfer to the electrode). Diverse community structures are such that the top 10
most predominant species will occupy >90% of the total biofilm population and it is
these main resident cells that will most determine the activity behaviour of the
MFC. For MFC using carbon veil anodes in continuous flow, the anodic chamber is a
good example of a perfusion matrix biofilm. Two important measurements which
are rarely mentioned in the MFC field and yet have a critical effect on MFC
performance are total biofilm population (biofilm yield) of attached cells within
the matrix around the electrode, and the numbers of cells released by the biofilm
to leave the anodic chamber as perfusate output. From these measurements, the
growth rate of the biofilm cells can be calculated and related to power output.

REFERENCES

- Aelterman P, Rabaey K, Pham TH, Boon N, Verstraete W. Continuous electricity


generation at high voltages and currents using stacked microbial fuel cells.
Environmental Science and Technology 2006; 40:3388–3394
- Cheng, S.; Liu, H.; Logan, B. E. (2006) Increased Power Generation in a
Continuous Flow MFC with Advective Flow through the Porous Anode and Reduced
Electrode Spacing. Environ. Sci. Technol. 40, 2426-2432
- Cohen, B. 1931. The Bacterial Culture as an Electrical Half-Cell. J. Bacteriol.,
Vol. 21, pp 18-19.
- Ieropoulos, I., Melhuish, C. and Greenman, J. 2003b: 'Artificial Metabolism:
Towards True Energetic Autonomy in Artificial Life', Proceedings of the 7th
European Conference in Artificial Life (ECAL 2003), Dortmund, Germany pp 792-
799
- Ieropoulos, I., Melhuish, C., Greenman, J. and Horsfield, I. 2005c. Artificial
symbiosis: Towards a robot-microbe partnership. Proceedings of the Towards
Autonomous Robotic Systems (TAROS '05) Conference, London, UK, pp 89-93.
- Ieropoulos, I., Greenman, J. and Melhuish, C. (2008) Microbial fuel cells based on
carbon veil electrodes: Stack configuration and scalability. Int. J. Energy Res.
32(13), 1228-1240. DOI: 10.1002/er.1419
- Melhuish, C., Ieropoulos, I., Greenman, J. and Horsfield, I. (2006) Energetically
autonomous robots: Food for thought. Auton Robot 21(3), 187-198.
- Potter, M., C. (1912) Electrical effects accompanying the Decomposition of
Organic Compounds. In Proc. Royal Soc. 84(Part B), 260-276.
- Rabaey, K.; et al. (2003) A Microbial Fuel Cell Capable of Converting Glucose to
Electricity at High Rate and Efficiency. Biotechnol. Lett. 25, 1531-1535.
- Wilkinson, S. 2000b. “Gastronome” – A Pioneering Food Powered Mobile Robot.
Proceedings of the 2000 IASTED Int. Conference on Robotics and Applications,
Paper # 318-037.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 43

CATHODES FOR SEDIMENT MICROBIAL FUEL CELLS

JAN B.A. ARENDS, ARNOUT D’HAESE, NICO BOON, WILLY VERSTRAETE

Laboratory of Microbial Ecology and Technology (LabMET) Ghent University, Faculty of


Bioscience Engineering, Coupure links 653, 9000 Gent, Belgium
Corresponding author email: Jan.Arends@Ugent.be

INTRODUCTION

Plant microbial fuel cells (P-MFCs), and more general sediment microbial fuel cells,
make use of electrogenic metabolism in anoxic soils and sediments. Electricigenic
micro organisms are able to respire organic carbon from the soil or sediment with
an electrode as electron acceptor. The power output of Plant MFCs is defined by a
vast number of factors among others; coulombic and energetic efficiency of the
anodic and cathodic reactions, mass and charge transport to and from the
electrodes and conductivity of the liquid and sediment. In this work the focus is on
the cathode. In a conventional sediment system an oxygen reducing cathode is
placed in the above water layer. In this new concept a drain tube was filled with
granules and placed in the sediment surrounded by the anode electrode granules.
This concept is compared with a conventional floating cathode and with a cathode
made of a tubular cation exchange membrane buried in the sediment.

MATERIALS AND METHODS

Two reactors of 8 L working volume were used. The bottom part was filled with
graphite granules (anode) and a tube filled with the same granules (cathode). The
tube in reactor A consisted of a commercial available drain tube 5 cm outer
diameter with cloth to prevent clogging. Reactor B had a 5 cm diameter tube made
of Ultrex cation exchange membrane (Clauwaert et al., 2007). Above the electrode
compartment sand (4 L) without any organic material was placed. The reactor was
filled with tapwater till 2 L above the sand. Daily additions of organics
corresponding to 20 ton dry weight ha-1 yr-1 were made by means of a 1:1 mix of
starch and acetate. To create a downward flux, daily Chemical Oxygen Demand
additions were accompanied by the removal of 0,5- 1L from the bottom and
addition of fresh tapwater to the top. Both cathodes were continuously
recirculated at 1,6 L/h (Prominent). Due to the nature of the drain tube, the
outflow of the cathode in reactor A is a mixture of anolyte and catholyte whereas
in reactor B both electrolytes stayed separated.
COD was measured with standard kits (Hach Lange, Germany). Polarization curves,
potential monitoring and electrical connections were made as mentioned
previously (Clauwaert et al., 2007). To start, all reactor compartments were
inoculated where necessary with effluent from performing MFC’s present in the
lab.
Firstly, the bottom cathodes of reactor A and B were studied. Later, an additional
cathode floating on top of the water layer was added and used for the study of the
internal resistances.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 44

RESULTS AND DISCUSSION

Potential
During the application of daily COD dosages, potential in the reactor A with the
drain tube cathode varied considerably. This is due to the change of oxic to anoxic
conditions in the anode. The permeable cathode barrier was able to deliver oxygen
to the anode compartment to make it oxic. However upon addition of COD this
compartment became anoxic as witnessed by the rise in cell potential and the
decrease in anode potential. Apparently the oxygen reducing cathode was not so
much affected by the presence of COD. The potential in the reactor with the
membranous cathode was rather stable. This is due to the fact that the membrane
is not penetrable by oxygen (Figure 1).

Organics
Daily COD recovery was determined in different runs. Overall recovery of COD as
current was similar between both reactors. For the drain tube reactor 2,15% of the
daily added COD was recover over a 24 hour feeding cycle (3 cycles). For the
reactor with the membranous cathode, COD recovery as current was 2,26% over a
24 hour feeding cycle (3 cycles). Potentials can be seen in (Figure 1).

In the reactor with the membranous cathode a build up of COD was witnessed in
the anode compartment, suggesting that this reactor was less efficient in the
breakdown of COD. COD concentrations were 7,5 x as high as in the reactor with
the drain tube cathode (460 mg/L vs 60 mg/L). Due to the mixing of the anode
liquid with the aerobic cathode liquid in the drain tube setup, most of the COD is
also removed aerobically whereas the membranous cathode reactor has no other
means to remove COD than through current and methanogenesis. Methanogenic
rate of COD removal was however not determined.

The COD data in combination with the potential data suggest that the resistance at
this point it to high to remove all COD efficiently. Maximum power output was
17mW/m2 for the drain tube cathode and sustained power output was 10 mW/m2
for the membranous cathode.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 45

0,6
A
0,5
0,4
0,3
Ecell
0,2
E (V)

Ean
0,1
Ecath
0
-0,1 0 0,5 1 1,5 2 2,5 3

-0,2
-0,3 Days
0,6
0,5 B

0,4
0,3
0,2 Ecell
E (V)

0,1 Ean
0 Ecath
-0,1 0 0,5 1 1,5 2 2,5 3
-0,2
-0,3
Days

Figure 1. Potentials from the reactor with the drain tube cathode (A) and
membranous cathode (B). Electrode potentials are reported vs NHE. Rext =470 Ω for
both reactors.

Internal resistance
Via the current interrupt method the total internal resistance of the various
sediment MFC configurations was determined. The same top cathode was used in
both reactor A and B. Reactor B with the membranous bottom cathode had the
lowest internal resistance. Interestingly it had the highest internal resistance with
the top cathode. In general, the bottom cathode configuration had a lower internal
resistance. Also the anode contribution to the internal resistance lowered when a
bottom cathode was present. Overall, the cathode had in all configurations a larger
contribution to the internal resistance than the anode.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 46

2,50

2,00 Reactor A 
Bottom
Rint (Ωm2)

1,50 Reactor A 
Top
1,00 Reactor B 
Bottom
0,50 Reactor B 
Top
0,00
Total Anode  Cathode
Figure 2. Internal resistances calculated via the current interrupt method (15 min
stabilization). The various internal resistances were calculated by Rint=E/J (E in
Volt, J in A/m2).

PERSPECTIVES

The configuration presented here is an extension of conventional thinking on


sediment MFC’s. However this configuration needs to be more rigorously explored
in terms of maximum power output and COD removal. Also the interaction with
plants needs more study. There are indications that active aeration can be omitted
while not hampering power output but this needs to be verified with lower
resistances and higher organic loading rates.

This configuration of a sediment MFC can be applied in greenhouses were drainage


and aeration of the rhizosphere are necessary for good plant health. Another less
high-tech application lies in a constructed wetland where the electrode
configuration can be applied as a BOD sensor, possibly as a safety to prevent BOD
spillage and as source of energy recovery (next to the energy contained in the
harvest plants) from the incoming (waste) stream.

REFERENCES

Clauwaert, P., Van der Ha, D., Boon, N., Verbeken, K., Verhaege, M., Rabaey,
K., Verstraete, W. 2007. Open air biocathode enables effective electricity
generation with microbial fuel cells. Environ. Sci. Technol., 41, 7564-7569.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 47

ANODE MATERIALS FOR SEDIMENT MICROBIAL FUEL


CELLS

JAN B.A. ARENDS1, EVELYNE BLONDEEL1, STEVE TENNISON2, NICO BOON1,


WILLY VERSTRAETE1
1
Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Faculty of
Bioscience Engineering, Coupure links 653, 9000 Gent, Belgium.
2
MAST Carbon International Ltd. Guildford Surrey GU3 2AF, United Kingdom.
Corresponding author email: Jan.Arends@Ugent.be

INTRODUCTION

Plant microbial fuel cells, and more general sediment microbial fuel cells, make
use of electricigenic metabolism in anoxic soils and sediments. Electricigenic
microorganisms are able to respire organic carbon from the soil or sediment with
an electrode as final electron acceptor. The power output of Plant MFCs is defined
by a vast number of factors among others; coulombic and energetic efficiency of
the anodic and cathodic reactions, mass and charge transport to and from the
electrodes and conductivity of the liquid and sediment. In this work the focus is on
the anode where current generation starts. To test the applicability of various
materials, microcosms with sediment MFCs were used.

MATERIALS AND METHODS

Glass jars with a volume of 200 ml were used for these studies. Various anode
materials (various available materials), see figure 1, were mixed with a layer of
sand. Cloth and felt materials had a projected area of 16 cm2 and only 1 layer,
interwoven with a graphite rod for electrical connection, was placed in the anode
compartment. Granular material was mixed 1:1 volume ratio with sand and a
graphite rod was introduced in the middle of the layer, at the same position as the
cloth materials. The same volume of sand was placed on top of the anode, which
can be considered a membrane, followed by an equivalent volume of liquid.
Modified M9 medium with 13 g/L NaCl was used in all tests. To start 20 mM
sodiumacetate was added to the medium in the anode compartment with 5 %
effluent of an acetate oxidizing anode. Cell potential (500 ohms) and anode
potential were measured continuously (Agilent benchlink datalogger).

RESULTS AND DISCUSSION

Materials
For all materials the start-up time before an increase in cell potential could be
seen was about 5 days. The exception to this was the granular material that was
already present in the anode from which the inoculum was obtained. This took
about two days to start. Total coulombs liberated were calculated from the
recorded potential over the resistor. This number is a good measure for how well a
particular material is able to accept and conduct electrons. From this measure it
can be seen that the larger granules (1-5 mm) were able to transfer the most
coulombs. The next best material can be considered all the cloth or felt materials
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 48

tested. The smaller granules were the worst performing material. Two fibre
materials cluster between the smaller granules. This is probably due to a bad
electrical connection between the material and the electrical circuit.

Cloth #13
G G F C F C C F F F G F G G G C

Granules #355
Granules #500
Granules #355
Felt 3,18 mm thick
Granules #500
Felt activated
Felt Carbonised
Felt Carbonised
Cloth #13
Cloth #20
Felt activated
Cloth #20
Felt 3,18 mm thick
Granules 1-5 mm
Granules 1-5 mm

0 20 40 60 80 100
Cumulative Coulombs transferred

Figure 1. Cumulative coulombs transferred after 8 days of incubation. C denotes


cloth material, F denotes felt material, G denotes a granular material.

Ratio Anode vs Soil mater


The commercial felt and granules of 1-5 mm were used for further study on the
ratio of material needed vs the amount of soil mater. The materials were mixed
with sand in different ratios to search for the best composition of the anode
compartment. Four different ratios were examined. The first setup didn’t have
any anode materials at the anode (only the graphite rod itself) and served as a
control. As for the granules, in the second setup 33% of the volume of the anode
was made out of granules, the third setup consisted of 67% and the fourth anode
compartment had a 100% volume of granules. With regard to the felt, the second
setup had an anode compartment made out of 14 %-vol felt, the third setup 28 %-
vol and the fourth 42 %-vol. To compare the different compositions the current
output per cross-section of anode was measured. All experimental setups were in
duplicate.

Granules
Granules were first monitored without any added carbon source to see the effect
of any electron donor present in the sand, granules or other materials. After 10
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 49

days acetate was added as the only carbon source at different spots in the anode.
The current output resulting from this addition can be seen in figure 2A.
2A 16 0%
14
0%
Current density J [A/m2]

12
33%
10
33%
8
67%
6
67%
4
2 100%

0 100%
0 2 4 6 8 10 12 14 16 18 20
2B Days
16
14
0%
Current density J [A/m2]

12
0%
10
14%
8 14%
6 28%
4 28%
2 42%

0 42%
0 2 4 6 8 10 12 14 16 18 20
Days
Figure 2. Current densities produced by granular (2A) and felt material (2B) in
different ratios in the soil. Arrow indicates feeding with acetate.

Clearly granule volume ratios 67% & 100% outperformed 0% & 33%. From these
results it can be seen that a 67% ratio of granules gives approximately the same
current output as 100% granules in the soil.

Felt
With regard to the felt, there was no clear distinction between the various ratios of
felt in the soil (figure 2 B). The felt did not reach a current output as high as the
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 50

granular material but when studying the current per % mixing, felt is the better
choice (table 1). This is possibly due to the better interconnectivity of felt as
compared to the granules.

Table 1. Current densities J [mA/m2] produced by granular and felt material in


different ratios in the soil after the 2nd addition of acetate. N.m. : not measured due
to equipment failure.
Days after 2nd feeding
Granules % 0 3 6
0 0,07 0,29 1,54
0 0,00 0,00 0,02
33 0,02 0,07 0,05
33 0,05 1,24 1,50
67 1,20 7,68 11,79
n.m.
67 1,27 4,41
100 0,36 4,94 12,54
100 1,91 3,80 14,78
Felt % 0 2 6
0 0,03 0,01 0,03
0 0,26 0,17 0,16
14 1,33 6,20 10,15
14 0,79 5,52 6,44
24 3,38 5,95 6,33
24 0,60 8,17 8,24
42 2,82 7,56 8,39
42 0,03 0,32 0,34

For the electrode/soil ratio test, anode potentials were also monitored. The
increase in cell potential, was mainly due to a decrease in anode potential to
values expected for anaerobic acetate oxidation ~ 250 mV vs NHE.

PERSPECTIVES

The study of various materials is necessary for their potential use at larger scale
systems with less control than a reactor environment. Granular material may have
an advantage when applying this to larger areas i.e. it can be ‘sown’. However, a
large amount needs to be applied to ensure electrical contact between most
granules. Felt materials might be a bit harder to install but have an overall better
electrical contact in the soil.
This work has provided a relative easy method to screen various materials for their
suitability as electrode materials for use in sediment fuel cell systems. The
methods described here will be used to further study various materials.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 51

AUTOTROPHIC NITROUS OXIDE REMOVAL IN


BIOELECTROCHEMICAL SYSTEMS
JOACHIM DESLOOVER1, SEBASTIA PUIG 2, WILLY VERSTRAETE1 AND NICO
BOON1.
1
Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Coupure Links
653, B-9000 Ghent, Belgium,
2
Laboratory of Chemical and Environmental Engineering (LEQUIA-UdG), Institute of the
Environment, University of Girona, Campus Montilivi s/n, Facultat de Ciències,
E-17071 Girona, Spain
Corresponding author E-mail: Joachim.Desloover@ugent.be

INTRODUCTION & OBJECTIVES

Bioelectrochemical systems (BESs) are devices where the oxidation of an electron


donor at the anode is coupled with the reduction of an electron acceptor at the
cathode, using bacteria to catalyze one or both reactions (Rabaey and Rozendal,
2010). Recently, it has been shown that biocathodes can perform autotrophic
denitrification by using the electrons and protons supplied by a bioanode
(Clauwaert et al., 2007; Puig et al., 2011; Virdis et al., 2008). The final step in the
denitrification pathway is the reduction of nitrous oxide (N2O) to dinitrogen gas
(N2) and was, until now, never thoroughly investigated in denitrifying biocathodes.
N2O is also an important greenhouse gas with an impact of about 298 times stronger
than that of carbon dioxide (Solomon et al., 2007), and is also considered as a
significant ozone-depleting substance (Ravishankara et al., 2009). Moreover,
anthropogenic sources of N2O represent at this moment almost 40% of the total N2O
emission (Reay et al., 2007).
This study investigates the N2O removal in a BES with a denitrifying biocathode.
This is the first proof of principle for BES technology as a possible technique for the
biological removal of N2O emissions from anthropogenic point sources.

MATERIALS & METHODS

A stack-type BES was used comprising an anodic and cathodic compartment (0.12 L
compartment), separated by a cation exchange membrane. Both compartments
were filled with granular graphite, resulting in a net anodic (NAC) and net cathodic
(NCC) compartment of 0.06 L each. The anode was always fed continuously with
acetate at a loading rate of 2.39 kg COD m-3 NAC d-1. The cathode was operated in
batch by adding a certain volume of 100% N2O in the headspace of the recirculation
vessel. N2O was the sole electron acceptor present.

RESULTS & DISCUSSION

The cathode of the BES was switched to a batch-fed operation mode with N2O after
a 30-day stable period of feeding the denitrifying biocathode continuously with
nitrate (loading rate: 0.209 ± 0.001 kg NO3-N m-3 NCC d-1; removal efficiency: 100
± 0%). The removal of N2O in the BES, current production and cathode potential in
function of time are presented in Fig. 1. The total nitrogen removal rate clearly
followed the same trend as the current production and the cathode potential.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 52

During the first 2.5h, a steady nitrogen removal rate of 4.57 ± 0.15 mg N h-1 (R2 =
0.9931) or 1.83 ± 0.01 kg N2O-N m-3 NCC d-1 and a maximum current production and
cathode potential of 7.3 mA (122 A m-3 NCC) and -0.14 V vs SHE were observed,
respectively. From that moment on, a gradual decrease of the nitrogen removal
rate, current production and cathode potential could be seen, until all N2O was
depleted. The obtained removal rate and current production were in the same
range as the values reported for denitrifying BESs with other electron acceptors
(nitrate or nitrite), when normalized to the amount of electrons needed to
complete the denitrification reaction to N2 (Clauwaert et al., 2007; Puig et al.,
2011; Virdis et al., 2008). The cathodic coulombic efficiency amounted 99%,
indicating that all electrons generated and used for current production where
recovered for the biological reduction of N2O to N2. Furthermore, no ammonium
(NH4+), nitrate (NO3-) and nitrite (NO2-) were detected in the cathodic liquid.
Control experiments (data not shown) revealed that no N2O was removed both
during open circuit operation (no passing through of electrons from anode to
cathode) and during operation with an abiotic autoclaved cathode. This
demonstrated that electrons derived from the anodic oxidation reaction of acetate
where the sole electron donors present in the cathode, and that denitrifying
microorganisms catalyzed the reduction reaction of N2O to N2.

Figure 1. Amount of N2O (total, gas and liquid), current production and cathode
potential in function of time in the BES.

The results obtained in this study demonstrated that a denitrifying biocathode is


capable to reduce N2O as the sole electron acceptor present. Furthermore, the
proof of principle is shown for BES technology as a possible biological treatment
technique for N2O emissions from anthropogenic point sources.

ACKNOWLEDGEMENTS

Joachim Desloover is recipient of a PhD grant from the Institute for the Promotion
of Innovation by Science and Technology in Flanders (IWT-Vlaanderen, SB-091144).
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 53

Sebastià Puig acknowledges the University of Girona (mobility grant 2010), Befesa
Agua S.A (CENIT-E TEcoAgua) and the Spanish Government (CONSOLIDER-CSD2007-
00055) for the financial support.

REFERENCES

Rabaey, K. and Rozendal, R.A. (2010) Microbial electrosynthesis - revisiting the


electrical route for microbial production. Nature Reviews Microbiology 8(10), 706-
716.

Clauwaert, P., Rabaey, K., Aelterman, P., De Schamphelaire, L., Ham, T.H.,
Boeckx, P., Boon, N. and Verstraete, W. (2007) Biological denitrification in
microbial fuel cells. Environmental Science & Technology 41(9), 3354-3360.

Puig, S., Serra, M., Vilar-Sanz, A., Cabre, M., Baneras, L., Colprim, J. and
Balaguer, M.D. (2011) Autotrophic nitrite removal in the cathode of microbial fuel
cells. Bioresource Technology (DOI: 10.1016/ j.biortech.2010.12.100).

Virdis, B., Rabaey, K., Yuan, Z. and Keller, J. (2008) Microbial fuel cells for
simultaneous carbon and nitrogen removal. Water Research 42(12), 3013-3024.

Solomon, S., Qin, D., Manning, M., Chen, Z., Marquis, M., Averyt, K.B., Tignor, M.
and Miller, H.L. (2007) Climate Change 2007: The physical science basis, Cambridge
University Press, Cambridge.

Ravishankara, A.R., Daniel, J.S. and Portmann, R.W. (2009) Nitrous Oxide (N2O):
The Dominant Ozone-Depleting Substance Emitted in the 21st Century. Science
326(5949), 123-125.

Reay, D., Hewitt, C., Smith, K. and Grace, J. (2007) Greenhous Gas Sinks, CAB
International, Oxfordshire, UK.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 54
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 55

YEAR ROUND PERFORMANCE OF THE FLAT-PLATE


PLANT-MICROBIAL FUEL CELL
M. HELDER, D.P.B.T.B. STRIK, H.V.M. HAMELERS, C.J.N. BUISMAN

Sub-department of Environmental Technology, Wageningen University, Bomenweg 2,


6703 HD, Wageningen, the Netherlands
Corresponding author E-mail: marjolein.helder@wur.nl

INTRODUCTION AND OBJECTIVES

Recent research into the Plant Microbial Fuel Cell (P-MFC) has shown that internal
resistances of the system are high and limit current and power density. Most
important resistance as measured by Timmers et al. was transport resistance [1].
The flat-plate P-MFC is a new design P-MFC in which distance between anode and
cathode is small and thus transport resistance can be reduced. Another aspect that
could improve power output of the P-MFC is the use of a plant-growth medium that
not only benefits plant-growth but does not hamper electricity production either.
In this research the flat-plate P-MFC was tested to see if we could limit internal
resistances, improve current and power density and relate electricity production to
root growth. Different plant growth media were tested to improve power output
further.

MATERIALS AND METHODS

The system was built as in figure 1 [2]. Both anode and cathode were split in three
sections to be able to measure electricity production on different depths in the
system. The system was operated for 365 days with different plant growth media.

A
1 2 3 A

B
1 2 3 B

C
1 2 3 C

a b

Figure 1. [2] Anode and cathode schematic overview in which 2a: anode
compartment with coding of anode-sections and sample points with: A, B, C =
separated parts of anode, 1, 2, 3 = sample spots in anode compartment. 2b with
flow-through channel and separated cathode sections indicated with A, B and C.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 56

RESULTS AND DISCUSSION

Internal resistances in the flat-plate system were only 0.101 Ωm2 [2], 4.5% of the
internal resistance as found by Timmers et al. [1]. Transport resistance was almost
zero. The low internal resistance led to a maximum power density of 440 mW/m2
and a maximum current density of 1632 mA/m2 geometric planting area [2]. This
power density is twice as high as maximally achieved in a P-MFC until now [3, 4].
The current density is more than 7 times higher than previously achieved current
densities [3]. The flat-plate PMFC was operated for a full year with Spartina anglica
with different growth media. Best results were achieved with a nitrate-less,
ammonium-rich plant-growth medium (see figure 2) [5]. After four months
electricity production started and an average current and power production of 169
mA/m2 and 37 mW/m2 were achieved during 8 months of electricity production.

1 2 3 1 3* 3 4 3 4 3
250
A
B
C
200

150
I (mA/m )
2

100

50

0
0 50 100 150 200 250 300 350
Time (days)

Figure 3. Electricity production in sections A, B and C of the P-MFC during 365 days
of experiment. Indicated with dotted lines are the different plant growth media
that were used, in which 1 = modified Hoagland 50%, 2 = modified medium without
nitrate and with ammonium chloride, 3 = modified medium with ammonium
bicarbonate. Indicated with 3* is the period in which medium 3 was added but the
system was not flushed with medium 3. 4= modified medium without sulphate.

During the year electricity production evolved from the top part of the system
gradually to the middle and the bottom of the system (see figure 2), consistent
with root growth in top, middle and bottom section.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 57

8000

7000

6000
Total leaflength (cm)

5000

4000

3000

2000

1000

0
0 50 100 150 200 250 300 350 400
Time (days)

Figure 3. Plant growth in total leaflength during 1 year of experiment

Above ground biomass in these 8 months grew from 43 to 305 stems and from 654
cm to 6791 cm total leaflength (figure 3), corresponding to 90.95 kJ when
converted into electricity. Total electricity production from below ground biomass
was 3760 J over the whole year, which is 4.1% of the electricity that could be
generated from above ground biomass.

REFERENCES

1. Timmers, R. A., Strik, D.P.B.T.B., Hamelers, H.V.M., Buisman, C.J.N., Long


term performance of a plant microbial fuel cell with Spartina anglica. Applied
Microbiology and Biotechnology 2010.

2. Helder, M., High power output with the flat-plate Plant-Microbial Fuel Cell.
submitted 2011.

3. Strik, D. P. B. T. B., Timmers, R.A., Helder, M., Steinbusch, K.J.J., Hamelers,


H.V.M., Buisman, C.J.N., Microbial solar cells: applying photosynthetic and
electrochemically active organisms. Trends in Biotechnology 2011.

4. Helder, M.; Strik, D. P. B. T. B.; Hamelers, H. V. M.; Kuhn, A. J.; Blok, C.;
Buisman, C. J. N., Concurrent bio-electricity and biomass production in three
Plant-Microbial Fuel Cells using Spartina anglica, Arundinella anomala and Arundo
donax. Bioresource Technology 2010, 101, (10), 3541-3547.

5. Helder, M., Strik, D.P.B.T.B., Timmers, R.A., Hamelers, H.V.M., Buisman,


C.J.N., Improved medium for the Plant Microbial Fuel Cell. in preparation 2011.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 58
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 59

DEHALOGENATION OF TRICHLOROETHYLENE IN
MICROBIAL ELECTROLYSIS CELLS WITH BIOGENIC
PALLADIUM NANOPARTICLES

T. HENNEBEL, B. DE GUSSEME, M. SOETAERT, S. DE CORTE, J. DE SLOOVER,


W. VERSTRAETE & N. BOON

Laboratory for Microbial Ecology and Technology, Ghent University, Coupure Links 653, 9000
Gent, Belgium
Corresponding author E-mail: Tom.Hennebel@ugent.be

INTRODUCTION

Nanopalladium catalysts can be synthesized by the precipitation of palladium (Pd)


on the surface of bacteria leading to the production of biogenic Pd(0) nanoparticles
(bio-Pd) (De Windt et al., 2005). Bio-Pd has been reported to catalyze efficiently
the dehalogenation of the groundwater contaminant trichloroethylene (TCE)
(Hennebel et al., 2009). In this conversion, bio-Pd was used as catalyst. Thus an
external electron donor such as hydrogen gas is required. In large-scale
applications, the use of hydrogen gas can give rise to large costs and technical
difficulties. Microbial electrolysis cells (MECs) can be used for the production of
hydrogen gas, however this innovative technology was never considered to deliver
hydrogen for bio-Pd catalyzed dehalogenation reactions. In MECs, organic material
is oxidized by electrochemically active microorganisms at the anode. Subsequently,
the microorganisms transfer the electrons resulting from this oxidation to the
anode. Through an electrical circuit, the electrons are transported to the cathode,
where they are consumed for hydrogen formation (Mu et al., 2009). In this study,
the application of bio-Pd in the cathode of an MEC as a catalyst for pollutant
reduction was compared with an MEC without bio-Pd and tested in the
dehalogenation reaction of the groundwater pollutant TCE. Additionally, it was
tested if different amounts of bio-Pd at the cathode influenced the dehalogenation
reaction at the cathode.

MATERIALS AND METHODS

Stack-type MECs containing graphite were constructed according to Clauwaert et


al. (2009). As electrolyte for anode and cathode, a minimal medium, consisting of 6
g L-1 Na2HPO42H2O, 3 g L-1 KH2PO4, 0.1 g L-1 (NH4)2PO4, 0.1 g L-1 Ca3(PO4)2 and
0.5 mL L-1 of a tracemetal solution (Clauwaert et al., 2009) was used. After the
successful start-up of the MEC, the cathode medium was spiked with 100 mg L-1
TCE for the dehalogenation experiments.

RESULTS AND DISCUSSION

Bio-Pd was produced as described by De Windt et al. (2005) and coated on the
graphite of the cathode to enhance TCE dechlorination. Two different Pd
concentrations were applied, i.e. 1 and 5 mg Pd g-1 graphite, and the Cl- and
ethane formation were monitored over 24 hours (Figure 1). When applying a cell
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 60

voltage of -0.8 V, a clear difference between the varying concentrations could be


observed. One mg Pd g-1 graphite in the cathode did not result in an improved
formation of Cl- or ethane in comparison to non bio-Pd containing graphite.
Moreover, the Cl- concentration after 24 hours amounted to 32 ± 6 mg L-1 Cl-, which
was lower than the formation of 50 ± 3 mg L-1 Cl- in the case no bio-Pd was used.
The same trend was observed for ethane production. After 24 hours, 12 ± 6 % of
the added TCE could be recovered as ethane with 1 mg Pd g-1 graphite versus 17 ±
7 % in the case of catalyst-free granules. In contrary, coating the granules with 5
mg Pd g-1 graphite resulted in faster and higher formation of both Cl- and ethane.
After 2 hours, already 80 ± 30 mg L-1 Cl- was formed versus 11 ± 5 mg L-1 Cl- in the
case of not-coated granules. At the end of the experiment 100 ± 30 mg L-1 Cl- was
detected in the cathode (Figure 1a). The ethane formation was produced more
rapidly and to a larger extent as well. After 4 and 24 hours respectively, 18 ± 3 %
and 26 ± 4 % of the initially added TCE amount could be recovered as ethane
(Figure 1b).

In case of the 1 mg Pd g-1 graphite, the TCE removal deteriorated while using 5 mg
Pd g-1 graphite enhanced the dehalogenation process. However, batch tests pointed
out that the 1 mg Pd g-1 graphite coated granules were catalytically active for
hydrodechlorination reaction of TCE. It is hypothesized that 2 mechanisms can
occur at the same time: (1) electrochemical reduction of TCE and (2) catalytical
dehalogenation with bio-Pd as catalyst and electrochemically formed H2 as the
hydrogen donor. Coating the granules with bio-Pd might negatively affect the first
mechanism due to the bacterial matrix, which is electrically less conductive, but
can enhance the second mechanism. However, also the Pd concentration plays a
crucial role as only 5 mg Pd g-1 graphite resulted in higher removal rates. Hence,
opting for the second mechanism requires a certain threshold of bio-Pd catalyst
and a cathode potential that allows for H2 production.

In this study, -0.8 V was applied to obtain hydrogen at a cathodic potential of


about -1 V vs. SHE. However, in theory, an applied voltage of only -0.14 V and a
cathodic potential of -0.42 V (vs. SHE) is required for hydrogen production through
biocatalyzed electrolysis of acetate. For example, Rozendal et al. (2006) obtained
H2 production at an applied voltage of -0.5 V and expected to lower this amount to
–(0.3 - 0.4) V. In any case, biocatalyzed electrolysis achieves very low energy
requirements for hydrogen production while water electrolysis in practice operates
at applied voltages well above 1.6 V (Rasten et al., 2003). This means that
biocatalyzed electrolysis requires four (Rozendal et al., 2006) to two (this study)
times less external energy. The price of hydrogen produced through water
electrolysis is strongly dependent on the electricity price. The lower consumption
of electrical energy per unit of hydrogen is a strong advantage of biocatalyzed
electrolysis. This opens perspectives for the economical and durable production of
hydrogen for bio-Pd catalyzed reactions. The use of bioelectrochemically produced
hydrogen gas would decrease the costs of this process with a factor of at least 2-4.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 61

Figure 1. Influence of the presence of bio-Pd (0, 1 and 5 mg bio-Pd g-1 graphite)
coated onto cathode graphite granules on (a) chloride and (b) ethane formation
versus time. Every experiment was performed in triplicate and mean ± standard
deviation are shown.

REFERENCES

Clauwaert P., Desloover J., Shea C., Nerenberg R., Boon N. and Verstraete W.
2009. Enhanced nitrogen removal in bio-electrochemical systems by pH control.
Biotechnol. Lett. 31:1537-1543.

De Windt W., Aelterman P. and Verstraete W. 2005. Bioreductive deposition of


palladium (0) nanoparticles on Shewanella oneidensis with catalytic activity
towards reductive dechlorination of polychlorinated biphenyls. Environ. Microbiol.
7:314-325.

Hennebel T., Simoen H., De Windt W., Verloo M., Boon N. and Verstraete W. 2009.
Biocatalytic dechlorination of trichloroethylene with bio-Pd in a pilot-scale
membrane reactor. Biotechnol. Bioeng. 102:995-1002.

Mu Y., Rabaey K., Rozendal R.A., Yuan Z.G., and Keller. J. 2009. Decolorization of
azo dyes in bioelectrochemical systems. Environ. Sci. Technol. 43:5137-5143.

Rasten E., Hagen G., and Tunold. R. 2003. Electrocatalysis in water electrolysis
with solid polymer electrolyte. Electrochim. Acta 48:3945-3952.

Rozendal, R.A., Hamelers H.V.M., Euverink G.J.M., Metz S.J., and Buisman. C.J.N.
2006. Principle and perspectives of hydrogen production through biocatalyzed
electrolysis. Int. J. Hydrogen Energy 31:1632-40.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 62
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 63

BACTERIAL MUTALISM IN THE MOSSES ROOTS


APPLICABLE IN
BRYOPHYTA-MICROBIAL FUEL CELL
YOLINA HUBENOVA1, MARIO MITOV2

1 - Department of Biochemistry and Microbiology, “Paisii Hilendarski” University of Plovdiv,


4000 Plovdiv, 24 Tzar Asen str., Bulgaria
2 - Department of Chemistry, South-West University ”Neofit Rilski”, 2700 Blagoevgrad,
66 Ivan Mihajlov str., Bulgaria
Corresponding author E-mail: jolinahubenova@yahoo.com

INRODUCTION AND OBJECTIVES

The Microbial Fuel Cell (MFC) technology is rapidly developed the last few years. It
is based on the utilization of whole microorganism cells and the conversion of a
part of their biochemical energy into electrical one. The investigations of higher
organisms such as plants as a future bioenergy source in the so-called Plant
Microbial Fuel Cell, however, have just begun. Actually the Plant-MFC technology
re-creates on a small scale the fundamental norms of the circulation of substances
and energy in the nature. The energy and food balance is divided between
conversion of carbon dioxide into organic compounds by using the energy of the
sunlight and the photolysis with dioxygen (O2) molecule release. In the Calvin cycle
the carbon fixation and reduction of carbon dioxide in the plant leaves leads to
production of 3 to 6-carbon carbohydrates. These small molecular weight
carbohydrates can be released by the plant roots excretion and together with the
dying particles of the roots themselves assimilated by the soil microorganisms
(reducents) yielding protons and electrons. The electrons can be donated to the
anode and due to the arising potential difference, to the cathode of a MFC.
The bryophytes are often considered as the amphibians of the plant world. In this
connection, we supposed that they could be successfully applied in plant MFC.
First, because the mosses could well adapt to a constant quantity of water, which
is necessary for their existence and can serve as an anolyte in MFC at the same
time. The second important reason is that the plant MFC - mini ecosystem produces
itself the essential for optimal microbial proliferation nutritious substances. And
last but not least, such MFCs are almost zero cost systems.
The aim of this ab initio study is to investigate a new type plant MFC, called by us
Bryophyta-MFC (B-MFC). This technological approach is based on both natural
developed species interactions combined with the MFC requirements for harvesting
in situ produced bioenergy.

MATERIALS AND METHODS

A forest moss Dicranum montanum was taken from tree roots in Rodopi mountain
and placed in a single-chamber MFC. Graphite rods were used as electrodes. The
cultivation was carried out under semi-natural conditions in a batch mode and
forest soil taken from the natural habitat. The electrolyte, added periodically, was
only potable water. The experimental window was 70 days and ranged over two
seasons – summer and autumn. The natural cultivation temperature varied in a
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 64

wide range: 16–35 °C in the summer and between 5 and 25 °C in the autumn
season. The moss cultivation was accomplished under direct sunshine during the
first part of the day.
Open circuit voltage (OCV) as well as terminal voltage under a load resistance 1 kΩ
was measured three times per day. From latter, current density values were
calculated.

RESULTS AND DISCUSSION

The observed exponential growth of data (Fig.1) coincides with the


microorganisms’ development and redistribution of microorganisms’ species in the
closed mini-ecosystem as well as on the electrode surface.

800 60
700
50
600
2
Current density, mA/m

40
500
OCV, mV

400 30

300
20
200
10
100

0 0
0 10 20 40 50 60 70
Time, days

Figure 1. Open circuit voltage and current density, R=1kΩ.with time.

The relatively constant values obtained between the 20th and 45th day demonstrate
that microorganisms’ layer was in a steady state. The further OCV leap up to
800 mV could be associated with an exchange of the microorganism species and a
development of new species of putrefactive bacteria or adapting of the microbial
society to the periodic polarization of the system (Fig.2).

800

700 without load

600

500
Voltage, mV

400

300
load 1k Ω load interruption
200

100

0
0 5 10 15 20 25 30
Time, minutes

Figure 2. Voltage recovery of the system.


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 65

Observations connected with a new growth of fresh leaves of the moss together
with the return of the high MFC values after drop, indicate the symbiotic function
between plant and microorganisms and their benefits for obtaining of useful MFC
outputs. The measurements during nights showed another tendency for increase of
current densities values, which could be connected to the Calvin cycles processes
in the dark.

For the first time the possibility for utilization of mutual society Bryophyta-
microorganisms as a natural bioreactor in Microbial Fuel Cell (MFC) was proved.
The accent was done on the bioelectrochemical properties of the system and the
current values, which can be generated. A single-chamber MFC attains cell voltage
values between 550-790 mV and current density between 20-40 mA/m2 after initial
period of 25 days. The observed fluctuation of MFC output is connected with the
complex variation of many factors such as environmental temperature, humidity,
illumination, silt-colloid particles and structure of the soil as well as the
alternation of different species participating at the decomposition processes.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 66
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 67

MODIFIED ELECTRODES FOR MORE POWERFUL AND


SUSTAINABLE PLANT-MFCS
MATTHIEU PICOT, LAURE LAPINSONNIÈRE, FRÉDÉRIC BARRIÈRE

(1) Université de Rennes 1, CNRS, UMR 6226, Sciences Chimiques de Rennes,


35042 Rennes cedex
(2) Université Européenne de Bretagne
Corresponding author E-mail: matthieu.picot@univ-rennes1.fr

One way to increase the power output of this system is the modification of the
anode and/or the cathode using different approaches. In MFCs, electrodes are
usually made of carbon in the form of felt, fibres or granules. The surface of these
materials can be modified either chemically or electrochemically by different way
,including the reduction of aryl diazonium salts which is described below (Fig 1).[1]
In this report we are investigating the impact of different anode modifications on
the MFCs performance.

R (A) R (B) R (C) R


Electrochemical or
chemical reduction
NaNO2

HCl

NH2 N2 BF4 Carbon surface Carbon surface


Fig 1. Scheme of carbon surface modification by diazonium

Figure 1. Scheme of carbon surface modification by diazonium.

In situ generation of the diazonium salt by adding sodium nitrite in acid media from
a starting aryl amine
Electrochemical (by applying a potential) or chemical (by adding a chemical
reductant which is commonly H3PO2) reduction of the diazonium salt which forms
the aryl radical upon release of N2.
Reaction at the vicinity of the surface between the electrode and the radical

The reduction of aryl diazonium salts is a powerful and versatile technique of


surface modification. In fact, it is possible to use a wide range of “R” groups (Fig
1). In this study we focused on potential charged groups (depending or not on pH).
The chemical groups selected are presented in figure 2.

These modified anodes were implemented into “H” type MFCs. It was
demonstrated that the charge on the surface had an effect regarding the power
density of the system. In fact, positively charged surface exhibited the highest
power output (Fig 3.a). However the negatively charged surface exhibit a power
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 68

output lower than the unmodified surface. (Fig 3. b). In fact it is well known that
the surface of bacteria is negatively charged [2]. Hence, electrostatic interactions
with the surface favour or inhibit the development and the connexion with the
biofilm which can be one way to explain the power output obtained.

COO

NH2 CH2 P

At pH=7 pH independent

Figure 2. Schematic of the selected groups.

SEM experiments were carried out on surfaces which were previously modified by
the positively charged group (a) and on the surface modified by the negatively
charged group (b) when the surface was colonized by the biofilm. Regarding these
pictures we can notice that the electrode modified by the positively charged group
is continuously colonized by the biofilm. On the other hand the other electrode is
not fully colonized which can be a result of the repulsive interaction between the
negatively charged surface and the bacteria cell wall. These modified anodes can
now be easily be implemented into MFCs and Plant MFCs [3] and tested as
biological cathodes as well.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 69

unmodified anode
0,12 anode modified with the negatively charged group
0,11
0,10 3A
0,09
power density (W/m2)

0,08
0,07
0,06
0,05
0,04
0,03
0,02
0,01
0,00

0,0 0,1 0,2 0,3 0,4 0,5 0,6 0,7


U(V)

unmodifed anode
anode modified with positively charged group
0,12
0,11
3B
0,10
power density (W/m )

0,09
2

0,08
0,07
0,06
0,05
0,04
0,03
0,02
0,01
0,00
0,0 0,1 0,2 0,3 0,4 0,5 0,6 0,7
U(V)

Figure 3. Power density curves of MFCs implemented with modified/unmodified


anodes.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 70

4A 4B

Figure 4. SEM pictures of colonized carbon surfaces previously modified.

REFERENCES

[1] F. Barrière, A. J. Downard, Journal of Solid State Electrochemistry 12 (2008)


1231-1244

[2] A.Terada, A.Yuasa, T. Kushimoto, S. Tsuneda, A. Katakai,M. Tamada,


Microbiology-Sgm 152 (2006) 3575-3583

[3] D. P. B. T. B. Strik, H. V. M. Hamelers, J. F. H. Snel, C. J. N. Buisman,


International Journal of Energy Research 32 (2008) 870-876
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 71

COULOMBIC EFFICIENCY IN A PMFC, EFFECT OF


EXUDATES TYPE AND CONCENTRATION, AND OXYGEN
STEINBUSCH, K.J.J., STRIK, D.P.B.T.B., HAMELERS, H.V.M.

Sub-department of Environmental technology, Wageningen University,


Bomenweg 2, 6703 HD Wageningen, The Netherlands
Corresponding author E-mail: bert.hamelers@wur.nl

INTRODUCTION

Estimating the coulombic efficiency for a plant microbial fuel cell (MFC) is more
complicated than for a normal microbial fuel cell (PMFC) 1. While a MFC is
manually fed with a measurable amount of soluble COD, a PMFC is supplied with an
unknown amount of soluble and solid COD via rhizodeposition of a plant. The exact
carbon and electron flow from the plant into the rhizosphere and finally the anode
is difficult to determine without disturbing the root system. The final current
density of a PMFC is determined by the total carbon/electron flow of the plant into
the rhizosphere and is diminished by the competitive processes that convert the
flow to other products than current. The exudation differs per plant species, the
plant age and stage of development; and by environmental factors as culture
conditions, nutrient deficiency, pH, water availability, temperature etc., and
varies in chemical structure (Grayston et al., 1997). Rhizodeposition comprises five
different exudates varying from soluble COD as low-molecular weight carboxylates
to solid COD as mucilage. The percentage of the electron flux from the root that is
eventually converted to electrons by electrical active bacteria is referred to as the
coulombic efficiency.

How much coulomb is produced from the excreted organic material? In general we
can say that the five different exudates are composed of the photosynthesis
building block sucrose, or more simplified glucose. Sucrose is a main transport
molecule of capture carbon in plants. One molecule of glucose can be oxidized to
24 electrons, 6 CO2 and water. In case glucose would be exuded 24 electrons can
be derived from the sucrose oxidation by electrochemically active bacteria. When
glucose is metabolized inside the plant, electrons end up in the final electron
acceptor and smaller organic molecules including acetate which may be exuded. In
the latter case, less electrons are available for electricity production from the
originally produced glucose. The objective is to quantify coulombic efficiency from
different type of exudates and to quantify the coulombic losses caused by
competitive processes as microbial oxidation en diffusion by plant roots at the
bioanode of the PMFC.

MATERIALS AND METHOD

Root exudation of low-molecular weight exudates was simulated in 2D set-up of a


root connected to a MFC, in which exudates and oxygen entered the anode by
diffusion. The coulombic losses in an anode were investigated by measuring the
current production, and at different distances from root to the membrane the
anode potential, pH, and the oxygen and substrate concentration. The cell was
continuously fed with 100% Hoagland medium, in which N-NO3 was replaced by N-
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 72

NH4. Each 7-1010 days, the substrate of the medium was changed concentration;
First, 1, 5 and 10 mM acetate was added at an exudation rate of 43.5, 216 and 435
µM d-1; Second, citrate was added to the medium in similar concentrations; Third,
dissolved oxygen was added to the medium.

Figure 1. Experimental
xperimental setup of the MFC with a known carbon flux.
flux

Experimental work was supported by calculations of the maximal power output for
exudates with low molecular weight as mono-,
mono di- and tricarboxylic acids. The
methodology used to calculate the theoretical
theoretical potential work and power is
described by Logan et al. 2. The anode potential was calculated for different
exudate type at pH 4 and 7. Acid dissociation constant (Ka) was taken into account
for the calculation of the cell potential at different pH values.
values. The chosen
concentration of bicarbonate was 0,05 M 3.

RESULTS AND DISCUSSION

The max. power output not only depends on the type of exudate but also at the pH
at which the exudate is oxidized (Figure 2). The performance depends on the
amount of electrons
rons that are theoretical available for the electrochemical active
microorganisms. Therefore, plants species with a higher exudation rate of for
example lactate or citrate may give a higher power output and could therefore be
selected for further experimental
experimen set-ups.
ups. Moreover, growing conditions such as
low phosphate concentration may enhance the release of those carboxylic acids 4.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 73

0.040
pH=4 pH=7
0.035

0.030
Power output (mW)

0.025

0.020

0.015

0.010

0.005

0.000
e

te
e

e
e

te
te

e
te

te
at
at

at

at

at

at

at
ra

tra
va
na

ala
et
rm

ct

in
al

ar
ty
ru

alo
io
Ac

Ci
Ox
La

M
cc

m
Fo

Bu
Py
op

Su

Fu
M
Pr

Monocarboxylate Di/Tricarboxylate

Figure 2. Maximum power output by oxidation of carboxylic acids at pH 4 and 7 at


an exudation rate recalculated in 1mM glucose per g dry root day-1. Recalculation
of mol carboxylic acids per mol glucose allows a fair comparison of the final
amount of electrons that are produced per mol photosynthesized transport
molecule glucose.

Experimental work with acetate and citrate was performed to support this theory.
It was expected that the exudation of citrate yields higher power output than the
exudation of acetate due to their higher degree of reduction. Preliminary results
will be presented at the symposium. Knowing the effect of different exudates and
oxygen on the coulombic efficiency of a PMFC, the cell can be optimized towards a
higher current density by conditioning the plant, rhizosphere and bioanode.

REFERENCES

1 Strik, D. P. B. T. B. et al. Microbial solar cells: Applying photosynthetic and


electrochemically active organisms. Trends in Biotechnology 29, 41-49 (2011).

2 Logan, B. E. et al. Microbial Fuel Cells:  Methodology and Technology.


Environmental Science & Technology 40, 5181-5192, doi:10.1021/es0605016 (2006).

3 Hamelers, H. et al. New applications and performance of bioelectrochemical


systems. Applied Microbiology and Biotechnology 85, 1673-1685,
doi:10.1007/s00253-009-2357-1 (2010).

4 Schwarz, D. & Grosch, R. Influence of nutrient solution concentration and a


root pathogen (Pythium aphanidermatum) on tomato root growth and morphology.
Scientia Horticulturae 97, 109-120 (2003).
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 74
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 75

POLYMER DERIVED CARBONS AND THEIR


APPLICATION IN PMFC’S
S. TENNISON AND D. E. BROWN

MAST Carbon International Ltd


Guildford Surrey GU3 2AF, United Kingdom.
Corresponding author E-mail: steve.tennison@MastCarbon.co.uk

Carbon materials form an essential part of the plant based fuel cells where their
primary role is to collect the power generated at the root system and to carry this
to surface although they could also contribute to other factors. The primary role is
superficially similar to the role of carbon materials in more conventional PEM fuel
cells although in some respects it is more simple whilst in other ways it is more
complex:

Roles for Carbon Materials


PEM Cell Plant Microbial Fuel Cell
Dispersed carbon supports the active Microbial catalysts are at the interface
catalysts at the anode and cathode. between the roots and the current
High metal dispersion required to give collector, intrinsically highly dispersed.
high power densities an reduced costs The carbon could function as a support
for the microbial catalysts. Fouling of
the active surface can easily occur.
Required to carry the oxygen and Could play a role in transporting
hydrogen from the gas phase to the oxygen to the cathode, fuel at the
supported catalysts and to transport anode provided by exudate from the
produced water away from the root system. Carbon could contribute
electrode to the distribution of nutrients?
Catalysts in intimate contact with the Power is generated at the root tips
carbon but require a complex three which grow and move through the soil
phase gas-liquid- solid interface to layer complicating the contact
function between the active sites and the
current collector system
Machined graphite substrate Much lower power densities (1.5
distributes gas across the cell cross A/m2)reduce the resistance constraints
section but also collects the current. but still important. Problem is how to
Very high power densities (up to 1.5 achieve good conductivity in a dynamic
A/cm2) make very low bulk and root environment
interfacial resistances critical

The cost of the carbon materials must also be taken into account. In a compact
high power density PEM cell the carbon components can account for ~30% of the
cell cost (~30% for the Pt metal,~ 30% the for polymer electrolyte) and include
higher performance machined graphite components and highly graphitised high
surface area carbon blacks, perhaps 300 Euro/kW power produced. In the much
lower power density PMFC’s, where a much larger area would be required for an
equivalent power generation the cost of the collector carbon could easily become a
dominant factor. The cost of carbon materials can vary between a few cents/Kg
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 76

and several Euro/g for the more exotic nano-materials that are being produced
today which would clearly have little place in a practical PMFC.

There is however a need to develop a better understanding of the role of the


carbon materials that can ultimately be used to design more cost effective carbons
for this complex application. This is difficult to achieve using naturally derived
carbons where the heterogeneity of the pore structure and surface chemistry
makes analysis of performance difficult if not impossible. This can however be
achieved through the use of advanced polymer derived carbon materials. These can
be produced as granular materials, with controlled pore structures in the micro,
meso and macro pore ranges, which can provide insights into the role of pore
structure in supporting the microbial components and in transporting fuel and
nutrients (Figure 1). In these materials the available surface is essentially internal
to the particles whilst the resistance within the cell will be dominated by inter-
particulate resistance.

Figure 1. Internal structure of mesoporous carbon

Alternatively they can be produced as extended two dimensional fabric or felt


systems based on a range of polymer precursors including viscose, PAN, phenolic
resin or pitch. These all exhibit different internal nano structures, surface
chemistry and external surface structures but they also provide a high geometric
surface due to small size of the fibres (~10microns) (Figure 2) with improved
resistivity along the fibre axis.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 77

Figure 2. Viscose derived carbon fibre (5 microns).

For all of the carbon materials the resistivity can be modified by heat treatment
although the response to temperature varies with the precursor. The phenolic resin
derived carbons are essentially amorphous and show only limited reductions in
resistivity between 800 and 2000 C. By contrast, pitch derived fibres are defined as
“graphitising” and show massive structural reorganisation across this temperature
range with very significant reductions in resistivity. PAN and viscose sit between
these two extremes.

Carbons are also unique in the extent to which the surface chemistry can be
tailored without major changes to the pore structure or physical form. The surface
acidity can be tailored by controlled gas phase or liquid phase oxidation whilst the
bascicity can be changed by thermal processing or by treatment with amines. The
impact of this on the behaviour of the PMFC components is as yet unknown
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 78
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 79

RADIAL OXYGEN LOSS DECREASES AVAILABLE


SUBSTRATE FOR ELECTROCHEMICALLY ACTIVE
BACTERIA IN A PMFC
RUUD A. TIMMERS, DAVID P.B.T.B. STRIK, CRISTINA ARAMPATZOGLOU,
HUBERTUS V.M HAMELERS, CEES J.N. BUISMAN

Sub-department of Environmental Technology, Wageningen University, Bomenweg 2, 6703 HD,


Wageningen, the Netherlands.
Corresponding author E-mail: bert.hamelers@wur.nl

INTRODUCTION

In the P-MFC roots are the key element because roots supply electrochemical
active bacteria with substrate in the form of organic rhizodeposisits.
Rhizodeposition of LMW organic compounds is a continuous process driven by
diffusion and migration. Both diffusion and migration and give a baseline
rhizodeposition of LMW organic compounds up to 2·10-8 mol m-2 roots-1 [1]. In
anoxic environments plant roots also loose oxygen into the rhizosphere due to
formation of aerenchyma which form an intracellular air space. ROL can vary from
2.5—10-7 through 2.6—10-9 mol O2 m-2 roots-1 dependent on location on the root and
physiological status of the root [2]. Oxygen plays an important role in the energy
recovery efficiency of a MFC because it can cause parasitic currents and parasitic
biological processes [3]. Parasitic currents occur when oxygen is reduced to water
in the anode of the MFC; oxygen reduction consumes electrons and therefore
lowers the number of electrons measured as electrical current. Parasitic biological
processes occur when oxygen is used as final electron acceptor by bacteria in the
anode of the MFC. Aerobic bacteria that use oxygen as final electron acceptor
consume substrate without production of current. Consumption of substrate
without the production of electrical current reduces the amount of substrate for
the electrochemically active bacteria and therefore lowers the number of electrons
available for current generation.
ROL can result in a micro oxic zone around the root of up to 2.5 mm [4] in which
both parasitic currents and parasitic biological processes possibly take place.
First objective was to determine the presence of oxygen in the anode of the P-MFC.
Second objective was to propose a model for the effect of ROL on energy recovery
in the P-MFC. Our approach was to measure oxygen concentration profiles in a P-
MFC that generated electrical current.

MATERIALS AND METHODS

PMFC set-up
The anode electrode of the P-MFC consisted of 165 g of graphite granules with a
diameter between 1 mm and 2 mm (le Carbone, Wemmel Belgium) within a glass
cylinder (Schott, Duran), diameter 35 mm, and height 300 mm. The glass cylinder
had a sample point every 30 mm along each side of the cylinder, numbered 1 to 8.
To separate the anode from the cathode a membrane (cation exchange membrane,
fumasep®, FKB) was fixed at the bottom of the glass cylinder. The cathode
compartment consisted of PVC beaker, diameter 110 mm, in which a 6 mm thick
graphite felt (Coidan Graphite Products LTD, York, United Kingdom). Catholyte was
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 80

50 mM ferric cyanide (K3[Fe(CN)]6 solution, buffered with 8 mM potassium


phosphate buffer (pH 6.8). The electrical circuit was closed over an external
resistance of 900 Ω.

Oxygen measurements
To measure oxygen in the anode an optical needle type oxygen micro-sensor was
inserted in the sample points after which the optical fiber sensor was pushed out of
the needle into the anolyte. The optical needle type oxygen sensor was connected
to an OXY-4 micro, 4 channel fiber-optic oxygen meter (Presens Precision Sensing
GmbH, Germany).

RESULTS AND DISCUSSOIN

Oxygen present in anode of P-MFC


Oxygen concentration in anode of the PMFC varied between 0.00 mg L-1 and 8.92
mg L-1. For a current generating MFC, Oh et al. measured oxygen concentrations
suggest a threshold bulk oxygen concentration of 1 mg L-1 in the anode above which
no current is generated [5]. The relative high oxygen concentration in anode of the
P-MFC compared to oxygen concentrations in anode of the conventional MFC can be
explained by ROL which resulted in micro oxic zones around roots in anode of the
P-MFC.
Oxygen concentration decreased with an increase in distance in the graphite bed
(figure 1A) which is consistent natural sediments. However oxygen is measured in
the anode of the PMFC at depths about 100 times deeper than in natural
sediments. Intracellular air space and micro oxic zones could explain these oxygen
measurements because root density decreased with distance in the anode of the
PMFC (figure 1B). Higher root densities increased the probability of measurements
in intracellular air paces or into micro oxic zones.
Oxygen (mg/l)
0 5 10

A 0 B
Depth in graphite bed (m)

-0.035

-0.07

-0.105

-0.14
d 40
-0.175 d 37

Figure 1A. Oxygen concentration measured in the graphite bed anode of a PMFC
generating 0.18 mA m-2 surface. (at the 37th and 40th day of operation). 1B.
Picture of root distribution in the graphite bed anode of a PMFC, showing a
decrease of the presence of roots with an increase in depth.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 81

Model for effect of ROL on energy recovery


An aerobic biofilm is present on root surface of G. maxima [6, 7]. The combination
of oxygen and aerobic biofilm reduces the available substrate for electrochemically
active bacteria and thus the energy recovery of the PMFC.
Based on numbers in literature for ROL the width of the oxic zone can be modeled
calculated based on: a flat aerobic biofilm, no accumulation of oxygen in the
aerobic biofilm, ROL from the root to the aerobic biofilm is known, zero order
kinetics for oxygen consumption of aerobic, and pseudo steady state: growth of the
aerobic film does not change width of the oxic zone. The amount of available
substrate EAB depends on the width of the oxic zone and can be modeled based on:
a known baseline rhizodeposition of LMW organic compounds, monod kinetics for
consumption rate of substrate (by AB) is zero order. Figure 2 shows the schematic
presentation of the model. Based on numbers given in the introduction the
maximum available substrate for EAB is 1.4·10-8 mol m-2roots-1 which results in a
maximum energy recovery of 74%. The maximum available substrate is equivalent
to a current of 32 mA m-2root (based on 24 e mol-1substrate). In literature numbers
for the root area index (RAI) (m-2root m-2surface) vary between 5 through 79 m-
2
root m-2surface [8]. With the RAI and maximum current per m-2root the maximum
current per m-2surface can be estimated by multiplication of the maximum current
with RAI. The maximum current per square meter of surface varies between 160
through 2528 mA m-2surface. average current densities reported in literature, 32
mA m-2surface [9], 44 mA m-2surface [10], 141 mA m-2surface [11], and 214 mA m-
2
surface [11], and 4.6 mA m-2surface [12] are in the lower range or below the
estimated maximum which indicates the possibility for improvement by either
suppressing ROL and increasing the RAI.

oxic zone anoxic zone


Intracellular gas space

LMW rhizodeposits
EAB

e to cathode
AB

H + CO2
+

ROL

root electrode
Figure 2. Schematic representation of model for effect of ROL on energy recovery
of a PMFC
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 82

REFERENCES

1. Jones, D.L., 1998. Plant Soil. 205, 25-44.

2. Soukup, A, 2007. New Phytol. 173, 264-278.

3. Harnisch, F., H. ChemSusChem. 2, 921-926.

4. Holmer, M. Mar. Ecol. Prog. Ser. 225, 197-204.

5. Oh, S.E. Water Sci. Technol. 60, 1311-1317.

6. Kowalchuk, G.A. FEMS Microbiol. Ecol. 27, 339-350.

7. Münch, C. Water Sci. Technol. 56, 271-276.

8. Jackson, R.B., 1997. Proc Natl Acad Sci USA. 94, 7363-7366.

9. Strik, D.P.B.T.B., 2008. Int. J. Energy Res. 32, 870-876.

10. De Schamphelaire, L., 2008. Environ. Sci. Technol. 42, 3053-3058.

11. Timmers, R.A., 2010. Appl. Microbiol. Biotechnol. 86, 973-981.

12. Helder, M., 2010. Biores. Tech. 101, 3541-3547


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 83

D:
IMPLEMENTATION & IMPACT
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 84

KEYNOTE BY PROF. DR. LOUISE VET


Louise E.M. Vet is a professor of Evolutionary Ecology at Wageningen
University and director of the Netherlands Institute of Ecology (NIOO-
KNAW), the largest institute of the Royal Netherlands Academy of Arts and
Sciences. She is an ecologist with a broad interest in ecology and
evolution, working on multitrophic interactions. She is internationally well
known for her work on plant-parasitoid interactions. In 1999 she moved
from Wageningen University to the NIOO. Here she expanded her research
with a new group to initiate studies on the interaction between above and
belowground multitrophic systems. Her research involves chemical,
behavioural and molecular ecology of plants and insects in a multitrophic
and community context. The research ranges from fundamental to
strategic: from questions on the evolution of species traits and species
interactions within communities to the strategic development of
sustainable agro-ecosystems that are primarily based on the prevention of
pests and diseases (life-support function of biodiversity).

She is an elected member of the Royal Netherlands Academy of Arts and


Sciences and published more than 185 papers in refereed journals (h-index
40). Vet was awarded several international prizes (e.g. British Rank Prize
for Nutrition).
Vet serves on a diversity of national and international boards and
committees. She has a special interest in communicating the importance of
science in general, and specifically ecology, to the general public
(lectures, columns, media). In addition to her professional interest in high
quality ecological scientific research, her recent outreach focuses on
achieving a positive interaction between ecology and economy. Practice
what you preach: a new NIOO building is presently being built with the
ambition to become the most sustainable laboratory/office complex in the
Netherlands. Linked to these building activities Vet stimulates public-
private partnerships to encourage new eco-technological developments.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 85

LEARNING FROM NATURE: NEED, CHALLENGE AND


IMPLEMENTATION OF ECO-TECHNOLOGY
PROF. DR. LOUISE E.M. VET

Netherlands Institute of Ecology (NIOO-KNAW)


Corresponding author E-mail: L.Vet@nioo.knaw.nl

Global ecosystems are endangered by rapidly growing demands for food, fresh
water, timber, fiber and fuel. Biodiversity is lost at an alarming rate by expanding
economies that ignore environmental degradation. When we talk about crises it is
not a financial crisis we should worry about, but the loss of resources and life on
earth. There is an increasing need to change from a linear, resource destroying,
take-make-waste economy towards a circular economy. But crises create room for
innovations. Nature can teach us valuable lessons for the transition towards a
different economy. After all, our planet has functioned for more than 3 billion
years without us in a sustainable way. Ecologists can preach these wise lessons but
it is even better when they practice them. Sustainable innovations, inspired by
nature, are the promises and challenges of the future. In this lecture I will present
an overview of some eco-technological applications, including Plant Microbial Fuel
Cells, in our new sustainable building.

CLAUS EN KAAN ARCHITECTEN

A building that breathes life


Early January 2011, the Netherlands Institute of Ecology (NIOO), one of the
research institutes of the Royal Netherlands Academy of Arts and Sciences (KNAW),
has merged its activities from two of its locations into one new sustainable
building. The NIOO studies the effect of nature in all its many forms. It is,
therefore, only fitting that ecological processes and the dynamics of nature
themselves have influenced the design and construction of their new premises.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 86

Cradle to Cradle
Inspired by the Cradle to Cradle (C2C) principles the design and construction of the
building was taken one step further than most sustainable buildings built to date in
the Netherlands. Sustainability is generally measured by energy efficiency; the C2C
concept, however, poses new criteria. The question is not what can we do to limit
environmental damage, but rather how can we make a valuable contribution to the
surrounding environment? The C2C guiding principles are the closing of cycles, the
use of solar energy and the celebration of biodiversity. The designers were
instructed to keep as close to this philosophy as possible. The office cum laboratory
now serves as a testing ground for eco-technology systems, where innovation and
experimentation are given room to grow.

Process
Having clarified the design brief, the next stage was to tackle the construction
process. Which materials could be used, what kind of flooring, how to generate
energy, how to close the wastewater circuit, and use the residual heat? Efforts
towards energy efficiency cover two areas: reducing consumption and sustainable
production, both of which lead to a reduction in CO2 emissions. The building uses
presence detection and daylight regulation switching. LED lighting is being used
where possible. Furthermore, a hybrid ventilation system is being installed. The
design encourages natural ventilation and thermal migration through the walls.
Mechanical ventilation is only enabled based on CO2 detection.
A trial is being conducted with the company Suncycle to develop a new generation
of energy-producing solar cells. The solar collector in the form of a sphere is
cheaper and more efficient than traditional solutions and also provides heat. On
the NIOO site, water will be used to check the cooling of these solar cells, the
warm cooling water can then be used to heat greenhouses and bioreactors. A
collection of thermal solar panels stores the sun's heat using unique High
Temperature Storage (HTS). Besides the solar heat, the HTS also stores the excess
heat from the building and the greenhouses. At a depth of 300 meters, the
temperature storage is located in much deeper geological strata than ever before.
The depth allows excess heat produced during the summer to be stored for use the
following winter. This innovative pilot project application produces energy savings
of 70% to 80%. The stored heat is delivered through pipes in the floors to the
interior (concrete core activation). Energy savings will increase in the coming years
as new technologies like solar energy plants and solar cells are refined and applied
on a larger scale.

Material
Claus and Kaan Architects had to meet a number of stringent material
specifications. The building had to be people and environmentally friendly, made
from renewable raw materials and economically produced without any harmful
emissions. The hull is made of durable concrete without any artificial additives and
no sealant, solvents or such like were used in the process. Using materials such as
wood, glass, steel, flax, ground limestone and granular debris creates a
streamlined building with an open and natural appearance.

Water
The approach to recycling is most visible in the water circuits. The objective is to
purify the waste water so that it can be discharged locally. In connection with this,
the NIOO found a connection to the sewers unnecessary. Building permits,
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 87

however, do not allow this. So, while there is a sewer, NIOO would prefer not to
use it. There are three different water circuits: rain water, domestic water
(including water from laboratories) and waste water from the toilets. After
purification (see below) the streams flow into a helophyte filter. Helophytes are
aquatic plants such as reed and cattails, which remove contaminants from the
waste water, thus reducing the ecological impact. The purified water then flows
into a pond and the open ditches in the surrounding area.

Waste = food
A truly complete recycling process is one that generates no waste. In treating all
waste as food, you create an ecological system that mirrors those found in nature.
This is one of the main principles of the Cradle to Cradle philosophy. Based on this
principle, a system is being developed for the NIOO-KNAW building that retrieves
valuable nutrients from faeces (in collaboration with a commercial company, DeSah
BV and Wageningen University). The system begins with the toilet. Vacuum toilets,
a unique concept in an office building, use a minimal amount of water. The
biomass is then passed into a thermophylic fermenter, where part of it is converted
into biogas, thereby linking the sanitation system to the energy system. The final
stages of water purification will involve an alga cultivation system and a helophyte
filter. NIOO and WUR scientists study the ability of algae to purify water (human
pathogens, pharmaceutical rest products, metals). By harvesting the algae valuable
minerals such as phosphates are recovered to be used as agricultural fertilizers.

Biodiversity
The NIOO-KNAW building will have a green roof. That goes without saying given
that roof vegetation filters water and air and aids temperature control. However,
NIOO-KNAW is taking it one step further. Together with Wageningen University
research is being conducted into how different green roofs function and contribute
to sustaining the variety in species of plants and animals. The prior Ministry of
Agriculture supports this project in the interests of saving endangered species of
plants listed under the European Habitats Directive. The site is being constructed
in harmony with the surrounding environment, where biodiversity is encouraged in
a variety of ways. The site will offer prime conditions for scientists and companies
to experiment with technology that generates power from living plants!

Integrated approach
Materials, water, energy, waste, and vegetation: these are not separate entities.
The challenge lies in the integration. The NIOO-KNAW is not striving to be the first,
the best, the smartest or the most innovative in one specific aspect of
sustainability, but to integrate a range of aspects. This poses a complex challenge,
or course, but it also brings great rewards. Linking the sanitation system to the
energy system is a unique process and promotes the effective use of what was
previously billed as ‘waste’. The benefits of a green roof are being combined with
the principles of biodiversity. Residual heat is stored long-term for future use.
These are examples of eco-effective design where the focus is not only on making
efficient use of separate systems, but also on creating an effective link between
the systems and residual flows.

A building that breathes life


The NIOO-KNAW building will never be 'finished'. There will always be room for
improvement and experimentation in the future. The building mirrors the dynamics
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 88

found in nature and will continue to adapt to new understandings and new
technologies. To aid the development of eco-technology, NIOO-KNAW is
encouraging companies and scientists to implement their ideas and applications for
environmental sustainability and the Cradle to Cradle method. Plant Microbial Fuel
Cell technology is being implemented by Plant-e on the roof and further up scaling
will be encouraged. NIOO is looking forward to it!
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 89

HARNESSING SOLAR ENERGY BY BIO-PHOTOVOLTAIC


(BPV) DEVICES

PAOLO BOMBELLI1,2, ALISTAIR MCCORMICK1,2, ROBERT BRADLEY2,


KAMRAN YUNUS1, JAMES PHILIPS1, XANDER ANDERSON3, SONIA CRUZ3,
REBECCA THORNE4, NING GU4, ALISON SMITH3, DEREK BENDALL2, CHRIS
HOWE2, LAURIE PETER4 AND ADRIAN FISHER1.

1
Department of Chemical Engineering and Biotechnology, University of Cambridge, New
Museums Site, Pembroke Street, CB2 3RA.
2
Department of Biochemistry, University of Cambridge, Hopkins Building, Downing Site, CB2
1QW.
3
Department of Plant Sciences, University of Cambridge, Downing Site, CB2 3EA.
4
Department of Chemistry, University of Bath, Bath. BA2 7AY.
Corresponding author E-mail: ajm283@cam.ac.uk

INTRODUCTION & OBJECTIVES

There is an urgent need to develop renewable energy technologies to replace


depleting fossil fuel resources and provide a carbon neutral source of power. Solar
energy is an abundant resource that represents an attractive target to supplement
this requirement and the development of efficient solar cell systems to capture
even a small fraction of this enormous reserve is currently an important scientific
and engineering challenge. Nature has clearly demonstrated that it is possible to
harness solar energy through the process of photosynthesis, and it is estimated that
the Earth’s photosynthetic organisms convert over 10 times more energy per year
than current human energy consumption, albeit with a low energy conversion
efficiency (ca. 0.25%). A number or synthetic techniques have been developed to
try and emulate the photosynthetic process; the most successful of these are
traditional silicon solar cells, which are based on the photovoltaic effect. Unlike
photosynthetic organisms, synthetic solar cells are able to convert energy with a
high efficiency (ca. 10%). However, the technology is based on the use of
expensive, high purity semi-conductor materials. Some of the key benefits of using
biological materials are that the photosensitive components are significantly
cheaper than synthetic analogues, are self-assembled and, in some cases, are
capable of self-repair. An ideal solar energy technology would attain the high
energy conversion efficiency of synthetic systems whilst keeping the inherent
merits of a low-cost biological approach. Bio-photovoltaics (BPV) describes a novel
method for harnessing solar energy which combines the synthetic and biological
techniques to produce an economical device with low manufacturing costs,
excellent energy conversion efficiency that is virtually carbon-free.

PROSPECTIVE

A recently established multidisciplinary consortium of groups based in the


University of Cambridge has proposed to develop, test and optimise BPV systems
(Fig. 1) by exploiting a wide range of techniques, including electrochemistry,
microfabrication, chemical synthesis, molecular biology, proteomics and numerical
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 90

simulation. The poster will describe significant research milestones already


achieved such as:

1. Developing a prototype BPV device as a proof of concept;


2. Optimising
ing power-output
power performance through device design;
3. Testing a range of photosynthetic materials, including intact single-cell
single
microorganisms.

In order to develop this technique as viable commercial technology, future


research efforts will be based upon developing
developing a fundamental understanding of the
electrochemical and metabolic processes occurring within the device. This
knowledge will then be applied to the optimisation of both the synthetic and
biological device components. This poster describes the proposed
proposed future work in
more detail:

1. Investigation of cell-surface
cell surface electron transfer at the molecular level;
2. Development of bio-compatible,
bio high-conductivity
conductivity electrode material;
3. Modelling of device performance by numerical simulation;
4. Fabrication of a pilot-device.
pilo

RESULTS & DISCUSSION


Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 91

Figure 1. Principles of operation of a bio-photovoltaic (BPV) system. A


schematic illustration of the key components of a typical BPV device (a). Within
the anodic chamber(4) the photosynthetic material releases electrons as a direct
consequence of photosynthetic activity or/and as result of catabolic metabolism of
organic substrates (e.g. carbohydrates). An electron mediator(5) siphons these
electrons from the photosynthetic material to the anode(3), Simultaneously, protons
(H+) diffuse from the anodic chamber across a proton exchange membrane(7) to the
cathode. Electrons from the anode travel through a copper wire(8) and an external
load(9) to the cathode(5), where they are combined with oxygen and H+ to re-form
water. The upper clamp(2) allows light(1) to enter the anodic chamber(4). Holes in
the lower clamp(10) allow oxygen access to the cathode and water to escape. Power
response of photosynthetic material upon illumination (b). Thylakoid membranes
(fine line) or Synechocystis sp. PCC 6803 (bald black line) was adjusted to 40 nmol
Chl ml-1. Ferricyanide (5 mM) was used as anodic electron mediator. The anodic
chamber area (0.8 cm2) was illuminated with 50 W m-2 at 670nm (LED). In the dark-
phase (black bar), Synechocystis produced a significant amount of power that
presumably reflects consumption of organic compounds by oxidative metabolism.
Conversely, thylakoids did not show any significant activity in the dark. Upon
illumination (light grey bar), thylakoid power output increased significantly faster
compared to Synechocystis, suggesting rapid electron transport from thylakoids to
the electron mediator. Thylakoids reached a maximum power output after 30 min
of illumination, following which power declined, indicating membrane degradation.
Conversely, power output from Synechocystis rose gradually during the light and
remained stable over time.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 92
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 93

MODELLING THE PLANT MICROBIAL FUEL CELL


H.V.M. HAMELERS, R.A. TIMMERS, K.J.J. STEINBUSCH & D.P.B.T.B. STRIK

Sub-department Environmental Technology, Wageningen University


Bomenweg 2, 6703 HD Wageningen, The Netherlands
Corresponding author E-mail: Bert.hamelers@wur.nl

The Plant Microbial Fuel Cell (P-MFC) is a promising technology for generating
renewable electricity. The technology is based on the Microbial Fuel Cell (MFC) in
which organic matter is oxidized and electricity is produced at high efficiency. The
MFC consists of an anodic chamber and a cathodic compartment. The MFC is based
on the presence of the Electrochemically Active Bacteria (EAB) in the anodic
chamber. The EAB are capable of oxidizing organic matter while transferring
electrons to the electrode. At the cathode the electrons are used to reduce oxygen
to water. Electrical energy can be drawn off from the current flowing between the
anode and the cathode.
The P-MFC is characterized by the fact that a plant is supplying the organic matter
to the anode, without being harvested. This makes the P-MFC attractive because no
energy is lost in harvesting, planting and transport. Furthermore is the P-MFC easy
to integrate in the landscape elements as parks and green roofs.

The source of organic matter is the rhizodeposition occurring at the roots of the
plants. By planting the plants with its roots in the anodic chamber the
rhizodeposition becomes available to the EAB. This EAB can generate than
electricity from the rhizodeposition.
The performance of the P-MFC is determined by the interplay of a number of
processes like photosynthesis, root growth, rhizodeposition, oxygen leakage,
hydrolysis, methanogenesis, electrogenesis, matter transport, electron transport
and oxygen reduction, to name some of the most important. It would be attractive
to have a model that describes theses processes. With such a model more insight in
the interaction between the different processes can be obtained. This insight can
lead to directions for improvement of the P-MFC performance.

The poster will describe for the first time such a model and will identify directions
for further improvement. The model is dynamic with respect to plan and root
growth and is able to simulate a whole growth season. All other process are
simulated as steady-state processes as their associated time constants are much
smaller than the duration of a growth season.

The model simulates the flow of electrons that are available from the organic
matter that is produced during photosynthesis. These flows are schematically
depicted in figure 1. The electrons are transported towards the roots, where part
of the electrons are respired, part is used for production of new roots and part end
up as exudates or dead biomass. The electrons that are released as exudates or
from dead roots can be either (i) oxidized with oxygen released by the roots, (ii)
converted into methane or (iii) oxidized while electrons are transferred to the
anode. All these processes are catalyzed by different groups of micro-organisms.
The process rate of the different processes is based on the Monod kinetics of the
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 94

associated physiological groups as present in the rhizosphere. The MFC itself is


modelled using a simple Randles equivalent circuit model.

Photosynthesis Electron Flow in P-


P-MFC (high tech)

Living Root Dead Root

Oxic Rhizosphere Anoxic Rhizosphere

O2
C-bound e Load
CH4

Electron Cathode
Cation

Figure 1. Electron Flow in high-tech P-MFC

As an example we show the current production over a growth season in figure 2.


The current starts to increases after an initial lag phase of 35 days. This is a result
of the development of the plant and roots that are needed to produces sufficient
exudates. The current keeps increasing during the whole period. This is in part due
to the increase of the exudating root mass but also of the organic matter
originating from the dead roots. Over time the fraction of current that comes from
the roots increases at the expense of the current that originates from exudates.
This means that during start-up, exudates are important while later in the growth
season the dead roots become also important.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 95

Figure 2. Development of current density during the growth season (red line, left
Y-axis) and its origin. The fraction coming from exudates (blue dotted, right Y-axis)
and the fraction originating from dead roots (green dotted, right Y-axis) are shown.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 96
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 97

ENERGETIC PERFORMANCE OF MICROBIAL SOLAR


CELLS
DAVID P.B.T.B. STRIK, RUUD A. TIMMERS, MARJOLEIN HELDER, KIRSTEN
J.J. STEINBUSCH, HUBERTUS V.M HAMELERS, CEES J.N. BUISMAN

Sub-department of Environmental Technology, Wageningen University, Bomenweg 2, 6703 HD,


Wageningen, the Netherlands.
Corresponding author E-mail: david.strik@wur.nl

INTRODUCTION

Microbial solar cells (MSCs) harvest solar energy to produce electricity. MSCs use
higher plants or photoautotrophic microorganisms to utilize solar energy, and use
electrochemically active bacteria in a microbial fuel cell (MFC) to generate
electrical current 1. MSCs are categorized according to the way solar energy is
harvested and/or transferred to the microbial fuel cell. Plant microbial fuel cells
(Plant-MFCs) are systems in which the plant roots directly fuel the
electrochemically active bacteria 2-4. Other MSCs have phototrophic biofilms
grown on electrodes to produce electricity 5. Also, there are MSCs which use
externally produced biomass as electron donor for the MFC. These systems grow
algae in photobioreactors, harvest phyto- and zooplankton from the coastal marine
ecosystems or use land produced energy crops 6-8. Here we overview the energetic
expectations and performance of these systems.

POTENTIAL

Figure 1 shows the potential net energy production in relation to the available
energy from organic matter production, the energy input of the system and the
MFC energy recovery. The numbers in the chart represent the net energy yield in
mW/m2 for Western European conditions1. The potential net energy production for
MSCs fed with energy crops was estimated from: 5% net photosynthetic efficiency
(=organic matter production), a minimum 10% organic matter loss due to
rhizodeposition, 60% MFC energy recovery and an estimated 30% energy input due
to transport and crop processing which is typical the case with anaerobic digestion
of energy crops 9-12.

Figure 1 shows that the highest net energy yield can be expected from Plant-MFCs
and MSCs using energy crops. MSCs with photobioreactors may have a relative high
organic matter production, the net energy yield may be diminished by the high
energy input. Phototrophic biofilms have are relative low net energy yield despite
their low organic matter productivity, but due to their negligible energy input.
Coastal marine ecosystems have an even lower organic matter production (too
little to be visible in figure 1) and a negative energy yield due to the needed
energy input to transport the organic matter to the MFC. Since MSCs which use
photobioreactors or coastal marine ecosystems may require an energy input higher
than the energy output, these systems must reduce the energy input will net
electricity generation be possible.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 98

12,000
organic matter production
40% MFC energy loss
10,000
energy input
net energy yield (mW/m2)
8,000

6,000

4,000 3,150
(mW/m2)
Energy

2,025
2,000

61
0

Energy crops
Coastal marine
Photobioreactor
Phototrophic

Plant-MFC

-2
biofilm

-2,000

-4,000
-4,195

-6,000 Microbial Solar Cell


Figure 1. Energetic potential of Microbial Solar Cells.

PERFORMANCE

Under lab scale conditions the Plant-MFC achieves an average power of 165 mW/m2
at a current density of 412 mA/m 13. This is 5% of the theoretical expectation. Lab
scale experiments of MSCs with phototrophic biofilms (not including sediments)
achieve average power densities of 2 mW/m2 at a current density of 6 mA/m2 5
which is 3% of the theoretical outlook. MSCs with photobioreactors, coastal marine
ecosystems or energy crops still have a negative net energy yield if we count the
needed energy input and the lab scale MFC power production1, 8, 14. We note that
it is still difficult to asses more precisely the net energy production of MSCs since
the systems are under research. Furthermore, we did not include the energy
input/output during the compete life-cycle of the different MSCs, which is needed
get the complete picture of the energetic performances.

PERSPECTIVES

So far, the most promising MSC is the Plant-MFC since this system has the highest
estimated and achieved net electricity generation. MSCs challenges towards
improvement energy recovery are: (i) improving the energy flux from the
phototrophic organisms to the electrochemically active organisms; (ii) improving
the energy recovery of the fuel cell, and (iii) decrease the energy input of MSCs
with photobioreactors, coastal marine ecosystems or ex-situ produced energy
crops. More details on these measures are reviewed elsewhere 1.
Comm. Appl. Biol. Sci, Ghent University, 76/2, 2011 99

REFERENCES

1. Strik, D.P.B.T.B., et al. (2011) Microbial solar cells: applying photosynthetic and
electrochemically active organisms. Trends in Biotechnology 29, 41-49

2. Strik, D.P.B.T.B., et al. (2008) Green electricity production with living plants
and bacteria in a fuel cell. International Journal of Energy Research 32, 870-876

3. Helder, M., et al. (2010) Concurrent bio-electricity and biomass production in


three Plant-Microbial Fuel Cells using Spartina anglica, Arundinella anomala and
Arundo donax. Bioresource Technology 101, 3541-3547

4. Timmers, R.A., et al. (2010) Long-term performance of a plant microbial fuel


cell with Spartina anglica. Applied Microbiology and Biotechnology 86, 973-981

5. Strik, D.P.B.T.B., et al. (2010) Solar energy powered microbial fuel cell with a
reversible bioelectrode. Environmental Science and Technology 44, 532-537

6. Strik, D.P.B.T.B., et al. (2008) Renewable sustainable biocatalyzed electricity


production in a photosynthetic algal microbial fuel cell (PAMFC). Applied
Microbiology and Biotechnology 81, 659-668

7. Girguis, P.R., et al. (2010) Harnessing energy from marine productivity using
bioelectrochemical systems. Current Opinion in Biotechnology 21

8. Pant, D., et al. (2010) A review of the substrates used in microbial fuel cells
(MFCs) for sustainable energy production. Bioresource Technology 101, 1533-1543

9. Neumann, G. (2007) Root Exudates and Nutrient Cycling. In Soil Biology


(Marschner P. , R.Z., ed), Springer-Verlag

10. Berglund, M., and Börjesson, P. (2006) Assessment of energy performance in the
life-cycle of biogas production. Biomass and Bioenergy 30, 254-266

11. Ter Heijne, A., et al. (2007) Microbial fuel cell operation with continuous
biological ferrous iron oxidation of the catholyte. Environmental Science and
Technology 41, 4130-4134

12. Taiz, L., and Zeiger, E. (2006) Plant Physiology. Sinauer Associates Inc.

13. Helder, M., et al. (submitted) High power output with the flat-plate Plant-
Microbial Fuel Cell.

14. Dekker, A., et al. (2009) Analysis and improvement of a scaled-up and stacked
microbial fuel cell. Environmental Science and Technology 43, 9038-9042